CN109310660B - Therapeutic compositions and uses thereof - Google Patents

Abstract

The present invention provides medicaments, including anti-epithelial cancer compositions, comprising glyceride compounds and compositions comprising one or more of these compounds. The invention also provides methods of making and using these compositions, particularly for the treatment or prevention of epithelial cancers, such as skin cancers, gastrointestinal cancers, and skin diseases.

Description

Therapeutic compositions and uses thereof

Technical Field

The present invention relates to compositions for skin and intestinal health care and for the treatment and prevention of epithelial cancers, including skin cancer and gastrointestinal cancer. In particular, the present invention relates to anti-epithelial cancer compositions comprising one or more glycerides containing dihydroxy fatty acids. Skin and intestinal health care and anti-epithelial cancer compositions comprising one or more dihydroxy fatty acid compounds are particularly contemplated, as well as the use of these compositions in the treatment or prevention of skin diseases, digestive tract diseases, mucosal surface diseases and epithelial cancers, including gastrointestinal cancers, such as colorectal cancer, laryngeal cancer, cheek cancer and stomach cancer, and skin cancers, such as basal cell carcinoma, squamous cell carcinoma and melanoma.

Background Art

Epithelial cancers include some of the most common cancers in the world. Colorectal cancer is reported as the second and third most common cancer in women and men in developed countries, respectively. Colorectal cancer is more common in developed countries—with the highest rates in the United States, Australia, Europe, and New Zealand—and is up to 10 times more common than in developing countries. Although surgery can be effective, early detection is essential for a positive surgical outcome. Other treatments are primarily aimed at prolonging life and palliation, as the effectiveness of chemotherapy and radiation therapy in treating primary tumors or metastases outside the lymph nodes is currently controversial.

Laryngeal cancer, also called esophageal cancer, pharyngeal cancer, or throat cancer, includes tumors that develop in the tissue of the pharynx, nasopharynx, oropharynx, hypopharynx, larynx (voice box), or tonsils. Treatment for laryngeal cancer includes surgery, radiation therapy, or chemotherapy. Treatment for laryngeal cancer may damage the throat and may also change the way a person eats, breathes, and sleeps. Similar problems currently exist with cheek cancer treatment.

Gastric cancer ranks second in cancer-related deaths worldwide. Because there are no symptoms in the early stages of gastric cancer, diagnosis is often delayed. The only way to treat gastric cancer is to remove the stomach through surgery (gastrectomy). Chemotherapy and radiation therapy after surgery may increase the chance of cure.

The incidence of skin cancer is also quite high, with more than 3 million cases diagnosed each year in the United States alone. In Australia and New Zealand, the incidence of skin cancer is particularly high, up to four times higher than in the United States. Although surgery may be effective, early detection is essential for a positive surgical outcome. Other treatments are mainly for low-risk disease, as the effectiveness of chemotherapy and radiation therapy in treating primary tumors, especially malignant melanoma or extra-lymph node metastases, is currently controversial.

Therefore, there is a need for anti-epithelial cancer compositions, including compositions suitable for treating or preventing skin cancer and/or gastrointestinal cancer, as well as compositions that can support the maintenance of anti-epithelial cancer activity or enhance anti-epithelial cancer activity.

The object of the present invention is to provide an anti-epithelial cancer composition for the treatment or prevention of epithelial cancers (e.g. skin cancer, including basal cell carcinoma, squamous cell carcinoma and melanoma) and/or gastrointestinal cancers (e.g. colorectal cancer, laryngeal cancer, cheek cancer and gastric cancer), or at least to provide the public with a useful choice.

Summary of the invention

Thus, in a first aspect, the present invention relates to a method for treating or preventing epithelial cancer in an individual, the method comprising administering to the individual in need thereof an effective amount of a compound of formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein R ₂ , R ₃ , R ₄ and R ₅ are independently H or acetyl (CH ₃ CO-),

_R6 is H or _CH3 , _R7 is _CH3 or a C2-C6 saturated or unsaturated hydrocarbon, and

x and y are independently integers of 3 to 14,

Provided that when R ₆ is H, x+y is greater than 10 and less than or equal to 18, and when R ₆ is CH ₃ , x+y is greater than 9 and less than or equal to 17. In one embodiment, the present invention relates to a method for treating or preventing epithelial cancer in an individual, the method comprising administering to an individual in need thereof an effective amount of a compound selected from any one or more of the following compounds

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In one embodiment, the method comprises administering to an individual in need thereof a composition comprising an effective amount of at least one compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or any one or more compounds selected from the group consisting essentially of, or consisting of

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In one embodiment, the present invention relates to a method for treating or preventing skin cancer in an individual, the method comprising administering to an individual in need thereof an effective amount of a composition comprising at least one compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or any one or more compounds selected from the group consisting of, or consisting mainly of,

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

On the other hand, the present invention relates to a method for inhibiting epithelial tumor formation, epithelial tumor growth or epithelial tumor metastasis in an individual, the method comprising administering to an individual in need thereof an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or any one or more compounds selected from the group consisting of

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In one embodiment, the present invention relates to a method for inhibiting skin tumor formation, inhibiting skin tumor growth or inhibiting skin tumor metastasis in an individual, the method comprising administering to an individual in need thereof an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or any one or more compounds selected from the following group

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

Another aspect relates to a method for inducing apoptosis of one or more tumor epithelial cells in an individual, the method comprising administering to an individual in need thereof an effective amount of a composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a therapeutically effective amount of at least one compound selected from any one or more of the following groups:

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In one embodiment, the method is a method of inducing apoptosis of one or more tumorous skin cells in an individual, the method comprising administering to an individual in need thereof an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or any one or more compounds selected from the group consisting of

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In one embodiment, apoptosis refers to apoptosis of basal carcinoma cells. In another embodiment, apoptosis refers to apoptosis of squamous carcinoma cells. In yet another embodiment, apoptosis refers to apoptosis of melanoma cells.

In another embodiment, the method is a method of inducing apoptosis in one or more tumor gastrointestinal cancer cells in a subject.

For example, apoptosis refers to apoptosis of colorectal cancer cells, laryngeal cancer cells, cheek cancer cells, or gastric cancer cells.

Another aspect relates to a method for regulating the proliferation of one or more tumor epithelial cells in an individual, the method comprising administering to an individual in need thereof an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or any one or more compounds selected from the group consisting of

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In one embodiment, the invention is directed to a method of regulating the proliferation of one or more neoplastic skin cells in a subject.

For example, in one embodiment, the modulation refers to reduction. Therefore, the present invention relates to a method for reducing the proliferation of one or more tumorous skin cells in an individual, the method comprising administering to an individual in need thereof an effective amount of a compound selected from any one or more of the following groups: 3,8-dihydroxyeicosanoic acid, 1-(3,8-dihydroxyeicosanoyl) glycerol, 1-(3,8-dihydroxyeicosanoyl) 2-acetoxy glycerol, 1-(3,8-dihydroxyeicosanoyl) 3-acetoxy glycerol, 1-(3,8-dihydroxyeicosanoyl) 2,3-diacetoxy glycerol, 1-(3,8-diacetoxyeicosanoyl) 2,3-diacetoxy glycerol, 3,8-dihydroxyeicosanoic acid methyl ester, 3,8-dihydroxyheneicosanoic acid, 1-(3,8-dihydroxyheneicosanoyl) glycerol, 1-(3,8-dihydroxyheneicosanoyl) glycerol 1-(3,8-dihydroxyhenicosanoyl) 2-acetoxyglycerol, 1-(3,8-dihydroxyhenicosanoyl) 3-acetoxyglycerol, 1-(3,8-dihydroxyhenicosanoyl) 2,3-diacetoxyglycerol, 1-(3,8-diacetoxyhenicosanoyl) 2,3-diacetoxyglycerol, 3,8-dihydroxyhenicosanoic acid methyl ester, 3,8-dihydroxydocosanoic acid, 1-(3,8-dihydroxydocosanoyl) glycerol, 1-(3,8-dihydroxydocosanoyl) 2-acetoxyglycerol, 1-(3,8-dihydroxydocosanoyl) 3-acetoxyglycerol, 1-(3,8-dihydroxydocosanoyl) 2,3-diacetoxyglycerol, 1-(3,8-diacetoxydocosanoyl) 2,3-diacetoxyglycerol and 3,8-dihydroxydocosanoic acid methyl ester.

In one embodiment, the proliferation refers to the proliferation of basal carcinoma cells. In another embodiment, the proliferation refers to the proliferation of squamous carcinoma cells. In yet another embodiment, the proliferation refers to the proliferation of melanoma cells.

In one embodiment, the method is a method of regulating the proliferation of one or more tumor gastrointestinal cancer cells in a subject. For example, the proliferation refers to the proliferation of colorectal cancer cells, laryngeal cancer cells, cheek cancer cells or gastric cancer cells.

Therefore, the present invention relates to a method for reducing the proliferation of one or more tumor gastrointestinal cancer cells in an individual, the method comprising administering to an individual in need thereof an effective amount of a compound selected from any one or more of the following groups: 3,8-dihydroxyeicosanoic acid, 1-(3,8-dihydroxyeicosanoyl)glycerol, 1-(3,8-dihydroxyeicosanoyl)2-acetoxyglycerol, 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol, 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol, 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol, 3,8-dihydroxyeicosanoic acid methyl ester, 3,8-dihydroxyheneicosanoic acid, 1-(3,8-dihydroxyheneicosanoyl)glycerol, 1-(3,8-dihydroxyheneicosanoyl) 1-(3,8-dihydroxyhenicosanoyl) 2-acetoxyglycerol, 1-(3,8-dihydroxyhenicosanoyl) 3-acetoxyglycerol, 1-(3,8-dihydroxyhenicosanoyl) 2,3-diacetoxyglycerol, 1-(3,8-diacetoxyhenicosanoyl) 2,3-diacetoxyglycerol, 3,8-dihydroxyhenicosanoic acid methyl ester, 3,8-dihydroxydocosanoic acid, 1-(3,8-dihydroxydocosanoyl) glycerol, 1-(3,8-dihydroxydocosanoyl) 2-acetoxyglycerol, 1-(3,8-dihydroxydocosanoyl) 3-acetoxyglycerol, 1-(3,8-dihydroxydocosanoyl) 2,3-diacetoxyglycerol, 1-(3,8-diacetoxydocosanoyl) 2,3-diacetoxyglycerol and 3,8-dihydroxydocosanoic acid methyl ester.

Another aspect relates to a method for inducing apoptosis of one or more tumor epithelial cells or regulating the proliferation of one or more tumor epithelial cells, such as a method for regulating the proliferation of one or more tumor epithelial cells in vitro or in vitro, the method comprising contacting one or more cells with an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or any one or more compounds selected from the group consisting of

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In another aspect, the present invention relates to a composition for treating or preventing epithelial cancer in an individual, the composition comprising, or consisting essentially of, or consisting of a therapeutically effective amount of a compound of formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein R ₂ , R ₃ , R ₄ and R ₅ are independently H or acetyl (CH ₃ CO-),

_R6 is H or _CH3 , _R7 is _CH3 or a C2-C6 saturated or unsaturated hydrocarbon, and

x and y are independently integers ranging from 3 to 14,

Provided that when R ₆ is H, x+y is greater than 10 and less than or equal to 18, and when R ₆ is CH ₃ , x+y is greater than 9 and less than or equal to 17.

In one embodiment, x=4, R6=H and y=9, 10, 11, 12 or 13.

For example, x=4, R ₄ , R ₅ ; R ₆ =H; y=10, 11 or 12.

Therefore, in another aspect, the present invention relates to an anti-epithelial cancer composition, which comprises at least one compound defined in the present invention, or is mainly composed of it, or is composed of it, wherein the at least one compound comprises a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or any one or more compounds selected from the following group

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In another aspect, the present invention relates to a pharmaceutical composition comprising, or consisting essentially of, or consisting of at least one compound as defined herein, wherein the at least one compound comprises a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a compound selected from any one or more of the following groups

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

In one embodiment, the composition is used to treat or prevent epithelial cancer.

In one embodiment, the composition is used to maintain or improve skin health.

In one embodiment, the composition is used to maintain or improve intestinal health.

On the other hand, the present invention relates to a method for inhibiting epithelial tumor formation, preferably skin tumor formation, inhibiting epithelial tumor growth, preferably skin tumor growth, inhibiting epithelial tumor metastasis, preferably skin tumor metastasis, or treating or preventing epithelial cancer, preferably skin cancer, in an individual, the method comprising administering an effective amount of a composition alone, simultaneously or continuously, the composition comprising a therapeutically effective amount of at least one compound defined in the present invention, or consisting mainly of it, or consisting of it, the at least one compound comprising a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a therapeutically effective amount of at least one compound selected from any one or more of the following

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

On the other hand, it relates to a method for improving the responsiveness of an individual to treatment of epithelial cancer, comprising administering to the individual an effective amount of a composition comprising a therapeutically effective amount of a compound of formula (I), or a therapeutically effective amount of at least one compound selected from any one or more of compounds a) to u) of the present invention, or a pharmaceutically acceptable salt or solvate thereof, or a composition of the present invention.

In one embodiment, epithelial cancer treatment refers to gastrointestinal cancer treatment. For example, gastrointestinal cancer treatment refers to colorectal cancer treatment, laryngeal cancer treatment, cheek cancer treatment or stomach cancer treatment.

In one embodiment, the invention relates to a method of increasing responsiveness of a subject to a skin cancer treatment comprising administering to the subject a composition of the invention.

In one embodiment, the skin cancer treatment is basal cell carcinoma treatment. In another embodiment, the skin cancer treatment is squamous cell carcinoma treatment. In yet another embodiment, the skin cancer treatment is melanoma treatment.

On the other hand, it relates to a method for increasing the sensitivity of an individual to treatment of epithelial cancer, comprising administering to the individual an effective amount of a composition comprising a therapeutically effective amount of a compound of formula (I), or a therapeutically effective amount of at least one compound selected from any one or more of compounds a) to u) of the present invention, or a pharmaceutically acceptable salt or solvate thereof, or a composition of the present invention.

Another aspect relates to a method for resensitizing one or more epithelial cancer cells that are resistant to treatment, the method comprising administering an effective amount of a composition comprising a therapeutically effective amount of a compound of formula (I), or a therapeutically effective amount of at least one compound selected from any one or more of compounds a) to u) of the present invention, or a pharmaceutically acceptable salt or solvate thereof.

In one embodiment, the epithelial tumor is a gastrointestinal tumor. For example, the gastrointestinal tumor is a colorectal tumor, a laryngeal tumor, an oral tumor, or a stomach tumor.

In one embodiment, the invention relates to a method of increasing the sensitivity of a skin tumor in an individual to a cancer treatment, comprising administering to the individual a composition of the invention.

In one embodiment, the skin cancer is basal cell carcinoma. In one embodiment, the skin cancer is squamous cell carcinoma. In another embodiment, the skin tumor is melanoma.

On the other hand, the present invention relates to a method for resensitizing one or more epithelial cancer cells that are resistant to treatment, the method comprising administering an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition described in the present invention to one or more epithelial cancer cells.

In one embodiment, the epithelial cancer cell comprises a tumor in a subject.

In one embodiment, the present invention relates to a method for resensitizing one or more epithelial cancer cells, preferably skin cancer cells that are resistant to treatment, comprising administering an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition of the present invention to one or more epithelial cancer cells, preferably skin cancer cells.

In one embodiment, the skin cancer cells comprise a tumor in a subject.

The present invention also relates to a method for at least partially reversing the resistance of tumor cells in an individual suffering from epithelial cancer to epithelial cancer treatment, the method comprising administering to the individual an effective amount of a composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a therapeutically effective amount of at least one compound selected from any one or more of compounds a) to u) of the present invention or a pharmaceutically acceptable salt or solvate thereof, or a composition of the present invention.

In one embodiment, the method is a method of at least partially reversing resistance of tumor cells to cancer therapy in an individual suffering from skin cancer, the method comprising administering to the individual an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a composition described herein.

The present invention also relates to a method for fully or partially reversing the resistance of an epithelial cancer patient, preferably a skin cancer patient, to an epithelial cancer treatment, preferably a skin cancer treatment, the method comprising the step of administering to the patient an effective amount of a composition, the composition comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a therapeutically effective amount of at least one compound selected from any one or more of the compounds a) to u) of the present invention or a pharmaceutically acceptable salt or solvate thereof, or a composition of the present invention.

In another aspect, the present invention provides a method for resensitizing one or more tumors in an epithelial cancer patient, wherein the one or more tumors are resistant to epithelial cancer treatment, or are expected to be resistant to epithelial cancer treatment. The method comprises the step of administering to the patient an effective amount of a composition, wherein the composition comprises a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a therapeutically effective amount of at least one compound selected from any one or more of compounds a) to u) of the present invention or a pharmaceutically acceptable salt or solvate thereof.

In one embodiment, the method is a method of resensitizing one or more tumors of a skin cancer patient, wherein the one or more tumors are resistant to skin cancer treatment or are expected to be resistant to skin cancer treatment. The method comprises the step of administering to the patient an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof or a composition described herein.

In one embodiment, the tumor is resistant to chemotherapy.

In one embodiment, the individual is a human. In another aspect, the present invention relates to a method for improving skin health or intestinal health, comprising administering an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof or a composition of the present invention to an individual in need thereof.

In one embodiment, the composition is a synergistic therapeutic composition. In one embodiment, the composition has a synergistic therapeutic effect.

In one embodiment, the composition includes a compound of the invention and at least one other therapeutic agent, and the therapeutic agent produces a synergistic therapeutic effect that is greater than the effect of either agent alone or greater than the additive effect of either agent alone. For example, there is a greater effect on induction of apoptosis, survival or proliferation of epithelial cancer cells, such as skin cancer cell survival or proliferation, resensitization to treatment, treatment or prevention of epithelial cancers, such as skin cancer, or responsiveness of an individual or tumor to a treatment method. Without being bound by theory, the inventors believe that this enhanced effect may be due to the bioavailability of the composition, such as by improving water dispersibility. In one embodiment, the compound and at least one other therapeutic agent may be administered in combination or sequentially for epithelial cancer treatment, such as skin cancer treatment, with appropriate reduction or increase in dosage or time of administration.

In one embodiment, the composition, such as a synergistic therapeutic composition, includes at least one other compound or extract of poplar propolis. For example, the composition also includes at least one compound selected from pinocytin, CAPE, chrysin, pinocytin chalcone, galangin, benzyl caffeic acid, benzyl ferulic acid or caffeic acid.

Another aspect relates to at least one compound of the present invention, including a compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof, or a compound selected from any one or more of the following

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

Used to prepare the medicine or composition for the purpose of the present invention.

In one embodiment, it is used together with at least one other therapeutic agent to prepare the medicament or composition for the purpose of the present invention.

Another aspect relates to the compounds of the present invention, including compounds of formula (I) or pharmaceutically acceptable salts or solvates thereof, or compounds selected from any one or more of the following

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

For use in preparing a medicament or composition for the purposes of the present invention together with at least one other therapeutic agent, wherein the medicament or composition is formulated for separate, simultaneous or sequential administration of the at least one compound and the at least one other therapeutic agent.

In one embodiment, the medicament or composition comprises cyclodextrin. In one embodiment, the medicament or composition comprises one or more dihydroxy fatty acid compounds of the present invention, such as compounds of formula (I) complexed with cyclodextrin.

In another embodiment, the compound is dissolved in a solvent or solvent mixture such as ethanol, ethanol/water, propanol, propanol/water, isopropanol, isopropanol/water, ethyl acetate, a hydrocarbon solvent, an edible oil, or propylene glycol.

In one embodiment, the drug or composition is a drug or composition to which at least one compound selected from any one or more of the following groups has been added

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester,

Or a pharmaceutically acceptable salt or solvate of any one of a) to u).

On the other hand, it relates to a composition for inhibiting epithelial tumor formation, epithelial tumor growth, epithelial tumor metastasis or treating or preventing epithelial cancer in a human individual; inducing apoptosis of one or more tumor epithelial cells in a human individual; increasing the responsiveness of a human individual to an epithelial cancer treatment; increasing the sensitivity of an epithelial tumor in a human individual to an epithelial cancer treatment; resensitizing one or more epithelial cancer cells in a human individual that are resistant to treatment; at least partially reversing the resistance of tumor cells in a human individual suffering from epithelial cancer to an epithelial cancer treatment; fully or partially reversing the resistance of an epithelial cancer patient to an epithelial cancer treatment; or resensitizing one or more tumors in a human patient with epithelial cancer, wherein the one or more tumors are resistant to an epithelial cancer treatment, or are expected to be resistant to an epithelial cancer treatment.

In one embodiment, the composition is used to inhibit skin tumor formation, skin tumor growth, skin tumor metastasis, or treat or prevent skin cancer in a human subject; induce apoptosis of one or more tumor skin cells in a human subject; increase the responsiveness of a human subject to a skin cancer treatment; increase the sensitivity of a skin tumor in a human subject to a skin cancer treatment; re-sensitize one or more skin cancer cells in a human subject that are resistant to treatment; at least partially reverse the resistance of tumor cells in a human subject with skin cancer to a skin cancer treatment; fully or partially reverse the resistance of a skin cancer patient to a skin cancer treatment; or re-sensitize one or more tumors in a human patient with skin cancer, wherein the one or more tumors are resistant to a skin cancer treatment, or are expected to be resistant to a skin cancer treatment.

In one embodiment, the invention relates to compositions for treating or preventing skin cancer.

In one embodiment, the composition further comprises cyclodextrin.

In one embodiment, the composition further comprises propolis, such as poplar propolis.

In one embodiment, the composition further comprises a poplar extract, such as the poplar leaf or bud exudate described in the embodiments of the present invention.

In various embodiments, the composition includes propolis, such as poplar propolis, including propolis extracts or fractions, propolis resins, propolis resin extracts and/or poplar extracts, wherein the propolis, propolis resins, propolis resin extracts and/or poplar extracts are enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof.

In one embodiment, the composition further comprises propolis, propolis resin or propolis resin extract and cyclodextrin.

In one embodiment, the cyclodextrin is α-cyclodextrin.

In one embodiment, the cyclodextrin is β-cyclodextrin.

In one embodiment, the cyclodextrin is γ-cyclodextrin.

In one embodiment, the cyclodextrin is hydroxypropyl β-cyclodextrin.

In one embodiment, the cyclodextrin is hydroxypropyl gamma-cyclodextrin.

In one embodiment, the cyclodextrin is a mixture of cyclodextrins.

In one embodiment, the anti-skin cancer composition is an anti-basal cell carcinoma composition. In another embodiment, the anti-skin cancer composition is an anti-squamous cell carcinoma composition. In yet another embodiment, the anti-skin cancer composition is an anti-melanoma composition.

In another aspect, the present invention relates to a pharmaceutical composition comprising propolis, propolis resin or propolis resin extract enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or mainly consisting of propolis, propolis resin or propolis resin extract, or consisting of propolis, propolis resin or propolis resin extract.

In one embodiment, the composition comprises propolis, propolis resin or propolis resin extract and one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, for example, the composition comprises propolis, propolis resin or propolis resin extract, to which one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof are added. In another embodiment, the composition comprises propolis and/or poplar bud or leaf exudate, propolis resin or propolis resin extract fractions enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof.

In one embodiment, the propolis is poplar propolis.

In one embodiment, the composition further comprises cyclodextrin.

In one embodiment, the composition is used to maintain or improve skin health. Therefore, in one embodiment, the present invention relates to a pharmaceutical composition for maintaining or improving skin health, the composition comprising propolis, propolis resin or propolis resin extract and cyclodextrin enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or mainly consisting of propolis, propolis resin or propolis resin extract and cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and cyclodextrin.

In one embodiment, the propolis enriched with one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof is a propolis resin, such as a poplar propolis resin. In one embodiment, the resin is a propolis resin, such as a poplar propolis resin; and the cyclodextrin is α, β or γ-cyclodextrin.

In another aspect, the present invention relates to a method for treating or preventing skin cancer in an individual, a method for inhibiting skin tumor formation, inhibiting skin tumor growth or inhibiting skin tumor metastasis, a method for inducing apoptosis of one or more neoplastic skin cells in an individual, a method for regulating the proliferation of one or more neoplastic skin cells in an individual, a method for increasing the responsiveness of an individual to skin cancer treatment, a method for increasing the sensitivity of a skin tumor in an individual to skin cancer treatment, and a method for resensitizing one or more skin cancer cells that are resistant to treatment; the method comprises administering to an individual in need thereof or to the one or more cells an effective amount of a composition comprising propolis, propolis resin or propolis resin extract enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and cyclodextrin, or consisting mainly of propolis, propolis resin or propolis resin extract and cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and cyclodextrin.

In one embodiment, the apoptosis refers to apoptosis of skin tumor cells, such as basal carcinoma, squamous cell carcinoma or melanoma cells.

In one embodiment, the modulation refers to reduction. Therefore, the present invention relates to a method of reducing the proliferation of one or more neoplastic skin cells in an individual, the method comprising administering to an individual in need thereof an effective amount of a composition comprising propolis, propolis resin or propolis resin extract enriched with one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof and cyclodextrin, or consisting essentially of propolis, propolis resin or propolis resin extract and cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and cyclodextrin.

In one embodiment, the proliferation refers to proliferation of skin tumor cells, such as proliferation of basal carcinoma, squamous cell carcinoma or melanoma cells.

In one embodiment, the skin cancer cell comprises a tumor in an individual. In one embodiment, the skin cancer cell is a basal cell carcinoma cell. In another embodiment, the skin cancer cell is a squamous cell carcinoma cell. In yet another embodiment, the skin cancer cell is a melanoma cell.

The present invention also relates to a method of at least partially reversing resistance of tumor cells in an individual suffering from skin cancer to skin cancer treatment, the method comprising administering to the individual a composition comprising propolis, propolis resin or propolis resin extract enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and cyclodextrin, or consisting essentially of propolis, propolis resin or propolis resin extract and cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and cyclodextrin.

The present invention also relates to a method for fully or partially reversing resistance to skin cancer treatment in a skin cancer patient, the method comprising the step of administering to the patient a composition comprising propolis, propolis resin or propolis resin extract and cyclodextrin enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or consisting mainly of propolis, propolis resin or propolis resin extract and cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and cyclodextrin.

In another aspect, the present invention provides a method for resensitizing one or more tumors in a skin cancer patient, said tumor being resistant, predicted to be resistant or likely to develop resistance to skin cancer treatment, said method comprising the step of administering to said patient a composition comprising, or consisting essentially of, propolis, propolis resin or propolis resin extract and cyclodextrin, enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or consisting of propolis, propolis resin or propolis resin extract and cyclodextrin.

In one embodiment, the one or more tumors are resistant to skin cancer treatment, are predicted to be resistant, or are likely to become resistant due to increased activity of one or more procancerous cell survival signaling pathways in one or more tumors or in the patient, including increased activity in one or more AKT, JNK, or JAK/STAT signaling pathways, for example, increased activity in a sample from the patient (e.g., a tissue sample, tumor biopsy, or a blood or plasma sample).

In one embodiment, the present invention provides a method for inactivating or inhibiting one or more precancerous cell survival signaling pathways in the one or more tumors or in the patient. For example, the present invention relates to a method for inactivating or inhibiting one or more AKT, JNK or JAK/STAT signaling pathways in the one or more tumors.

In one embodiment, said one or more tumors are resistant, predicted to be resistant, or likely to develop resistance to skin cancer treatment due to increased activity of one or more of said AKT, JNK, or JAK/STAT signaling pathways within said tumor.

In one embodiment, the invention provides a method of preventing a tumor from developing resistance to a primary skin cancer treatment, wherein said resistance is mediated at least in part by increased activity of one or more of said AKT, JNK or JAK/STAT signaling pathways, for example, within said tumor.

In one embodiment, the tumor is resistant to chemotherapy.

In one embodiment, the skin cancer is basal cell carcinoma. In another embodiment, the skin cancer is squamous cell carcinoma. In yet another embodiment, the skin cancer is melanoma.

In one embodiment, the composition comprises, is mainly composed of, or is composed of propolis, propolis resin or propolis resin extract, α-cyclodextrin (e.g., poplar propolis, propolis resin or propolis resin extract) and cyclodextrin (e.g., poplar propolis and α-cyclodextrin) enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof.

In one embodiment, the composition comprises, is mainly composed of, or is composed of propolis, propolis resin or propolis resin extract, β-cyclodextrin (e.g., poplar propolis, propolis resin or propolis resin extract) and cyclodextrin (e.g., poplar propolis, propolis resin or propolis resin extract and β-cyclodextrin) enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof.

In one embodiment, the composition comprises, is mainly composed of, or is composed of propolis, propolis resin or propolis resin extract, γ-cyclodextrin (e.g., poplar propolis, propolis resin or propolis resin extract) and cyclodextrin (e.g., poplar propolis, propolis resin or propolis resin extract and γ-cyclodextrin) enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof.

On the other hand, the present invention provides a synergistic composition, comprising propolis, propolis resin or propolis resin extract rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and cyclodextrin. In one embodiment, the composition is a synergistic therapeutic composition. In one embodiment, the composition has a synergistic therapeutic effect.

In one embodiment, propolis, propolis resin or propolis resin extract and cyclodextrin enriched with one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof all have a synergistic therapeutic effect, which is greater than the effect of a single one or greater than the additive effect of a single one. For example, it has a greater effect on induction of apoptosis, survival or proliferation of skin cancer cells, resensitization of treatment, treatment or prevention of skin cancer, or responsiveness of an individual or tumor to a treatment method. In one embodiment, propolis, propolis resin or propolis resin extract and cyclodextrin enriched with one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof can be administered, co-administered or continuously administered to treat skin cancer, and the dosage or duration of the medication can be reduced or increased as appropriate.

Another aspect relates to propolis, propolis resin or propolis resin extract and cyclodextrin enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, which can be used to prepare the composition for the purpose of the present invention.

Another aspect relates to propolis, propolis resin or propolis resin extract and cyclodextrin enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, which can be used together with at least one other therapeutic agent to prepare a composition for the purpose of the present invention. In one embodiment, concentrated propolis, propolis resin or propolis resin extract and α-cyclodextrin are used. In one embodiment, concentrated propolis, propolis resin or propolis resin extract and β-cyclodextrin are used. In one embodiment, concentrated propolis and γ-cyclodextrin are used.

Another aspect of the present invention relates to a complex comprising propolis, propolis resin or propolis resin extract enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof and cyclodextrin, wherein the complex is used together with at least one other therapeutic agent to prepare a composition for the purpose of the present invention, wherein the composition is formulated so as to be administered separately, simultaneously or consecutively with propolis, propolis resin or propolis resin extract, cyclodextrin complex and at least one other therapeutic agent.

On the other hand, it relates to a composition for improving skin health, comprising propolis, propolis resin or propolis resin extract and cyclodextrin, which are rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or mainly consisting of propolis, propolis resin or propolis resin extract and cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and cyclodextrin. In one embodiment, the composition comprises propolis, propolis resin or propolis resin extract and α-cyclodextrin, which are rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or mainly consisting of propolis, propolis resin or propolis resin extract and α-cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and α-cyclodextrin. In one embodiment, the composition comprises propolis, propolis resin or propolis resin extract and γ-cyclodextrin enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or is mainly composed of propolis, propolis resin or propolis resin extract and γ-cyclodextrin, or is composed of propolis, propolis resin or propolis resin extract and γ-cyclodextrin.

In another aspect, the present invention relates to a composition for treating or preventing skin cancer in an individual, the composition comprising propolis, propolis resin or propolis resin extract enriched with one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and cyclodextrin, or consisting mainly of propolis, propolis resin or propolis resin extract and cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and cyclodextrin. In one embodiment, the composition comprises propolis, propolis resin or propolis resin extract enriched with one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and α-cyclodextrin, or consisting mainly of propolis, propolis resin or propolis resin extract and α-cyclodextrin, or consisting of propolis, propolis resin or propolis resin extract and α-cyclodextrin. In one embodiment, the composition comprises propolis, propolis resin or propolis resin extract and γ-cyclodextrin enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or is mainly composed of propolis, propolis resin or propolis resin extract and γ-cyclodextrin, or is composed of propolis, propolis resin or propolis resin extract and γ-cyclodextrin.

Another aspect relates to a product comprising one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and cyclodextrin, and optionally, one or more, two or more, or three or more other therapeutic agents for simultaneous, separate or sequential administration as a co-formulation according to the purposes of the present invention.

Another aspect relates to a composition for treating or preventing skin cancer.

One aspect relates to a method for separating or purifying a compound of formula (I) or a mixture of compounds from propolis or poplar, propolis resin or an extract or exudate thereof, comprising the following steps

Providing poplar, propolis, propolis resin or extracts or exudates thereof, and

Isolate or purify a compound or mixture of compounds from poplar, propolis, propolis resin or an extract or exudate thereof.

In various embodiments, the method includes one or more of the following non-limiting steps:

a) fractionating poplar, propolis, propolis resin or an extract or exudate thereof by chromatography (e.g. column chromatography, reverse phase chromatography, normal phase chromatography or supercritical fluid chromatography, and/or solvent separation and/or supercritical extraction) to produce one or more fractions comprising one or more compounds of formula (I),

b) fractionating poplar, propolis, propolis resin or an extract or exudate or one or more fractions thereof by preparative HPLC and/or polymeric resin separation to produce one or more fractions comprising one or more compounds of formula (I), and optionally,

c) further purifying one or more compounds of formula (I) by one or more components of step b) and/or step c).

In another embodiment, poplar, propolis, propolis resin or extract or exudate can be subjected to hydrolysis, methylation or acetylation reaction, and then subjected to one or more steps a), b) or c) to isolate and purify one or more compounds of formula (I).

Another aspect relates to markers of anti-epithelial cancer efficacy, including compounds of formula (I).

Another aspect relates to a method for evaluating the anti-epithelial cancer efficacy of a composition or product, comprising:

a) providing a sample of the composition or product, and

b) or

i) determining the presence or absence of one or more compounds of formula (I) in a sample, or

ii) measuring the amount and/or concentration of one or more compounds of formula (I) in the sample and determining whether the amount and/or concentration of the one or more compounds in the sample is equal to or greater than a reference amount of a known compound having an anti-epithelial cancer therapeutic effect, and optionally,

iii) Determining the biological activity of the compound and/or compositions containing the compound using in vitro or in vivo bioassays.

Another aspect relates to a method for evaluating the anti-epithelial cancer efficacy of a composition or product, comprising:

a) receiving information about the presence and/or amount of one or more compounds of formula (I) in a sample of a composition or product, and

b) determining whether the amount of each compound in the sample is equal to or greater than a reference amount of a known compound having anti-epithelial cancer efficacy.

Another aspect relates to a method for evaluating the anti-epithelial cancer efficacy of a composition or product, comprising:

a) determining the presence and/or amount of one or more compounds of formula (I) in a composition or product sample using gas, liquid or supercritical fluid chromatography, and

b) determining whether the amount of each compound in the sample is equal to or greater than a reference amount of a known compound having anti-epithelial cancer efficacy.

In various embodiments, the method of evaluating the anti-epithelial cancer efficacy of a composition or product comprises any one or more of the following steps:

a) fractionating the sample by solvent separation and/or supercritical extraction and/or chromatography (e.g. reverse phase chromatography) to produce one or more fractions enriched in one or more compounds of formula (I),

b) further fractionating the one or more components enriched in one or more compounds of formula (I) by one or more solvents or supercritical extraction or chromatography to produce one or more components enriched in one or more compounds of formula (I),

c) analyzing the highly concentrated fraction, for example, by one or more gas, liquid or supercritical chromatography, to determine the presence and/or amount of one or more compounds of formula (I) in the sample.

In various embodiments, the method of evaluating the anti-epithelial cancer efficacy of a composition or product comprises any one or more of the following steps:

a. fractionating the sample by chromatography (e.g., reverse phase chromatography) to generate one or more chromatographic components,

b. fractionating the sample or chromatographic fraction by preparative HPLC to produce one or more HPLC fractions,

c. Analyze the traces generated by preparative HPLC to determine if the compound is present and/or in what quantity in the sample, and/or

d. performing mass spectrometry analysis on the sample, one or more reverse phase chromatography fractions or one or more HPLC fractions to determine the presence and/or quantity of one or more compounds of formula (I) in the sample.

In one embodiment, the presence and/or amount of one or more compounds of formula (I) in a sample is determined by mass spectrometry, such as liquid chromatography mass spectrometry (LC-MS).

In one aspect, the present invention relates to a composition or product comprising one or more compounds of formula (I), wherein the amount and/or concentration of the one or more compounds of formula (I) is specified as indicated, such as but not limited to a product label, a certificate of analysis, a website, or promotional materials associated with the composition or product.

In various embodiments, the compound of formula (I) is selected from the group consisting of

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol,

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol,

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxydocosanoyl)propylene glycol,

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol, and

u) 3,8-dihydroxydocosanoic acid methyl ester.

In one embodiment, the composition or product is or comprises propolis, propolis resin or a propolis resin extract, for example propolis, propolis resin or a propolis resin extract comprising or enriched in one or more compounds of formula (I).

In one embodiment, the instructions associated with the composition or product are a label or package insert.

In one embodiment, the instruction sheet associated with the composition or product is a certificate of analysis (COA).

In one embodiment, the instructions associated with the composition or product are a website promoting the product.

In one embodiment, the instructions associated with the composition or product are a brochure promoting the product.

The following embodiments may relate to any of the above aspects.

In various embodiments, the epithelial cancer is epidermal cancer or epidermoid cancer.

In various embodiments, the epithelial cancer refers to gastrointestinal cancer, such as colorectal cancer, laryngeal cancer, esophageal cancer, cheek cancer, or gastric cancer. For example, in one embodiment, the colorectal cancer refers to colon adenocarcinoma. In another embodiment, the esophageal cancer refers to esophageal squamous cell carcinoma. In another embodiment, the gastric cancer refers to gastric tumor.

In other embodiments, the epithelial cancer is skin cancer, such as basal cell carcinoma, squamous cell carcinoma, or melanoma.

In various embodiments, the composition further comprises one or more of caffeic acid phenethyl ester (CAPE), caffeic acid, pinocytin, benzylcaffeic acid, benzylferulic acid, benzylisoferulic acid, chrysin, cinnamocaffeic acid, pinocytin chalcone, galangin, and pinocytinol.

In an exemplary embodiment, one or more of CAPE, caffeic acid, pinocysteine, benzylcaffeic acid, benzylferulic acid, isobenzylferulic acid, cinnamocaffeic acid, pinocysteine chalcone, chrysin, galangin and pinocysteine alcohol-3-acetate are added to the composition.

In one embodiment, the composition has a CAPE concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the composition has a pinocybin concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the composition has a galangin concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, and an effective range can be selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the composition has a chrysin concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the composition has a pinocybin concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the caffeic acid concentration of the composition is greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the composition has a benzylcaffeic acid concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the composition has a benzylferulic acid concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the composition has a cinnamocaffeic acid concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In one embodiment, the composition has a cinnamiferulic acid concentration of greater than about 1, 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 75, 80, 90, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, or 600 mg/g, with an effective range selected from any of these values (e.g., about 1 to about 5, about 1 to about 10, about 2 to about 20, about 5 to about 20, about 5 to about 25, about 10 to about 25, about 10 to about 40, about 15 to about 100, or about 20 to about 600 mg/g).

In various embodiments, the cyclodextrin is α-cyclodextrin, or the cyclodextrin is present as a combination of cyclodextrins including α-cyclodextrin.

In various embodiments, the cyclodextrin is β-cyclodextrin, or the cyclodextrin is present as a combination of cyclodextrins including β-cyclodextrin.

In various embodiments, the cyclodextrin is γ-cyclodextrin, or the cyclodextrin is present as a combination of cyclodextrins including γ-cyclodextrin.

In one embodiment, the cyclodextrin is a modified cyclodextrin, examples of which are shown in Stella He, Toxicological Pathology, Vol. 36, 2008, pp. 30-42.

In one embodiment, the modified cyclodextrin is hydroxypropyl beta cyclodextrin or hydroxypropyl gamma cyclodextrin.

In one embodiment, the propolis is present in the anti-epithelial cancer composition as a propolis extract or a component.

In one embodiment, the propolis contained in the anti-epithelial cancer composition does not contain wax. For example, the propolis has been dewaxed using an extraction method known in the art. Dewaxed propolis is generally known or referred to as "propolis resin".

In one embodiment, the propolis is "poplar-type" or "poplar" propolis. For example, "poplar-type" propolis is derived at least in part from bud and leaf exudates of one or more of poplar, birch, larch or willow.

In one embodiment, the composition comprises about 1.0%wt to about 99%wt of propolis, propolis resin or an extract of propolis resin enriched in one or more compounds of formula (I). In one embodiment, the composition comprises about 1.0%wt to about 99%wt of propolis resin enriched in one or more compounds of formula (I).

In various embodiments, the composition includes about 1% wt to about 99% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), about 1% wt to about 25% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), about 1% wt to about 30% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), about 1% wt to about 40% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), about 1% wt to about 50% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), about 5% wt to about 25% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), about 5% wt to about 10% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), and about 5% wt to about 25% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I). from about 5% wt to about 40% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), from about 5% wt to about 50% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), from about 5% wt to about 99% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), from about 10% wt to about 25% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), from about 10% wt to about 30% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), from about 10% wt to about 40% wt of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I), Propolis, propolis resin or propolis resin extracts containing about 10% wt to about 50% wt of a compound of formula (I), propolis, propolis resin or propolis resin extracts containing about 10% wt to about 99% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts containing about 15% wt to about 25% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts containing about 15% wt to about 30% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts containing about 15% wt to about 40% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts containing about 15% wt to about 50% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts containing about 15% wt to about 99% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts containing about 15% wt to about 99% wt of one or more compounds of formula (I), Propolis, propolis resin or propolis resin extracts containing about 20% wt to about 25% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts enriched in about 20% wt to about 30% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts enriched in about 20% wt to about 40% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts enriched in about 20% wt to about 50% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts enriched in about 20% wt to about 99% wt of one or more compounds of formula (I), propolis, propolis resin or propolis resin extracts enriched in about 25% wt of one or more compounds of formula (I), or propolis, propolis resin or propolis resin extracts enriched in about 30% wt of one or more compounds of formula (I).

In various embodiments, the composition comprises about 1% wt to about 99% wt of propolis resin enriched in one or more compounds of formula (I), about 1% wt to about 25% wt of propolis resin enriched in one or more compounds of formula (I), about 1% wt to about 30% wt of propolis resin enriched in one or more compounds of formula (I), about 1% wt to about 40% wt of propolis resin enriched in one or more compounds of formula (I), about 1% wt to about 50% wt of propolis resin enriched in one or more compounds of formula (I), about 5% wt to about 25% wt of propolis resin enriched in one or more compounds of formula (I), about 5% wt to about 30% wt of propolis resin, about 5% wt to about 40% wt of propolis resin enriched in one or more compounds of formula (I), about 5% wt to about 50% wt of propolis resin enriched in one or more compounds of formula (I), about 5% wt to about 99% wt of propolis resin enriched in one or more compounds of formula (I), about 10% wt to about 25% wt of propolis resin enriched in one or more compounds of formula (I), about 10% wt to about 30% wt of propolis resin enriched in one or more compounds of formula (I), about 10% wt to about 40% wt of propolis resin enriched in one or more compounds of formula (I), Propolis resin containing about 10% wt to about 50% wt of a compound of formula (I), propolis resin containing about 10% wt to about 99% wt of one or more compounds of formula (I), propolis resin containing about 15% wt to about 25% wt of one or more compounds of formula (I), propolis resin containing about 15% wt to about 30% wt of one or more compounds of formula (I), propolis resin containing about 15% wt to about 40% wt of one or more compounds of formula (I), propolis resin containing about 15% wt to about 50% wt of one or more compounds of formula (I), propolis containing about 15% wt to about 99% wt of one or more compounds of formula (I) Resin, propolis resin enriched in about 20%wt to about 25%wt of one or more compounds of formula (I), propolis resin enriched in about 20%wt to about 30%wt of one or more compounds of formula (I), propolis resin enriched in about 20%wt to about 40%wt of one or more compounds of formula (I), propolis resin enriched in about 20%wt to about 50%wt of one or more compounds of formula (I), propolis resin enriched in about 20%wt to about 99%wt of one or more compounds of formula (I), propolis resin enriched in about 25%wt of one or more compounds of formula (I), or propolis resin enriched in about 30%wt of one or more compounds of formula (I).

In one embodiment, the propolis, propolis resin or propolis resin extract in the composition is completely included in cyclodextrin.

In one embodiment, the molar ratio of propolis, propolis resin or propolis resin extract to cyclodextrin in the composition is no greater than about 1:1.

In one embodiment, the propolis, propolis resin or propolis resin extract is poplar propolis, propolis resin or propolis resin extract.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 3,8-dihydroxyeicosanoic acid. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxyeicosanoyl) glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxyeicosanoyl) 2-acetoxy glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxyeicosanoyl) 3-acetoxy glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxyeicosanoyl) 2,3-diacetoxy glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 3,8-dihydroxyeicosanoic acid methyl ester. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 3,8-dihydroxyheneicosanoic acid. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxyheneicosanoyl)glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxyhenedecanoyl) 3-acetoxyglycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxyhenedecanoyl) 2,3-diacetoxyglycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-diacetoxyhenedecanoyl) 2,3-diacetoxyglycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 3,8-dihydroxyhenedecanoic acid methyl ester. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 3,8-dihydroxybehenic acid. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxybehenoyl) glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxybehenoyl) 2-acetoxy glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxybehenoyl) 3-acetoxy glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-dihydroxybehenoyl) 2,3-diacetoxy glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract includes and/or is rich in 1-(3,8-diacetoxybehenoyl) 2,3-diacetoxy glycerol. In one embodiment, the poplar propolis, propolis resin or propolis resin extract comprises and/or is enriched in 3,8-dihydroxydocosanoic acid methyl ester.

In one embodiment, the poplar propolis has any one or more of the compounds a) to u) of the present invention in a concentration greater than about 1 mg/kg, about 1.5 mg/kg, about 2 mg/kg, about 2.5 mg/kg, about 3 mg/kg, about 3.5 mg/kg, about 4 mg/kg, about 4.5 mg/kg, about 5 mg/kg, about 5.5 mg/kg, about 6 mg/kg, about 7.5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20 mg/kg g, about 25 mg/kg, about 30 mg/kg, about 40 mg/kg, about 50 mg/kg, about 75 mg/kg, about 100 mg/kg, about 125 mg/kg, about 150 mg/kg, about 175 mg/kg, about 200 mg/kg, 250 mg/kg, about 300 mg/kg, about 350 mg/kg, about 400 mg/kg, about 450 mg/kg, about 500 mg/kg, about 550 mg/kg, or about 600 mg/kg.

In some embodiments, the poplar propolis, propolis resin or propolis resin extract has any two or more, three or more, four or more of the compounds a) to u) described herein, which is at least about 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 7.5, 10, 15, 20, 25, 30, 40, 50, 75, 100, 125, 150, 175, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 or 1000 mg/g; an effective range can be selected from any of these values (e.g., about 1 to about 400, about 1 to about 100, about 5 to about 1000, about 5 to about 500, about 5 to about 300, about 5 to about 100). For example, in some embodiments, the poplar propolis has 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol and 1-(3,8-dihydroxybehenosanoyl)3-acetoxyglycerol, or 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol, 1-(3,8-dihydroxybehenosanoyl)3-acetoxyglycerol and 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol, or 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol, 1-(3,8-dihydroxybehenosanoyl)3-acetoxyglycerol and 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol, and any one or more of compounds a) to c), e) to j) or l) to u) at a concentration of about 1 mg/g to 1000 mg/g.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g. g, about 25 mg/g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 3,8-dihydroxyeicosanoic acid.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 2 About 500 mg/g, about 550 mg/g, about 600 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxyeicosanoyl)glycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. / g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxyeicosanoyl) 2-acetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. / g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxyeicosanoyl) 2,3-diacetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. , about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g. , about 25 mg/g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of methyl 3,8-dihydroxyeicosanoate.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g. g, about 25 mg/g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 3,8-dihydroxyheneicosanoic acid.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 2 About 5 mg/g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxyheneicosanoyl)glycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. / g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxyheneicosanoyl) 2-acetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. / g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. , about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g, about 30 mg/g, about 35 mg/g, about 36 mg/g, about 37 mg/g, about 38 mg/g, about 39 mg/g, about 40 mg/g, about 41 mg/g, about 42 mg/g, about 43 mg/g, about 44 mg/g, about 45 mg/g, about 46 mg/g, about 47 mg/g, about 48 mg/g, about 49 mg/g, about 50 mg/g, about 51 mg/g, about 52 mg/g, about 53 mg/g, about 54 mg/g, about 56 mg/g, about 58 mg/g, about 59 mg/g, about 60 mg/g, about 61 mg/g, about 62 mg/g, about 63 mg/g, about 64 mg/g About 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g , about 25 mg/g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of methyl 3,8-dihydroxyheneicosanoate.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g. g, about 25 mg/g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 3,8-dihydroxydocosanoic acid.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 2 About 500 mg/g, about 550 mg/g, about 600 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxybehenanoyl)glycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. / g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxybehenoyl) 2-acetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. / g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxybehenoyl)3-acetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g. , about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-dihydroxybehenoyl) 2,3-diacetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g, about 25 mg/g, about 30 mg/g, about 35 mg/g, about 36 mg/g, about 37 mg/g, about 38 mg/g, about 39 mg/g, about 40 mg/g, about 41 mg/g, about 42 mg/g, about 43 mg/g, about 44 mg/g, about 45 mg/g, about 46 mg/g, about 47 mg/g, about 48 mg/g, about 49 mg/g, about 50 mg/g, about 51 mg/g, about 52 mg/g, about 53 mg/g, about 54 mg/g, about 56 mg/g, about 58 mg/g, about 59 mg/g, about 60 mg/g, about 61 mg/g, about 62 mg/g, about 63 mg/g, about 64 mg/g About 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of 1-(3,8-diacetoxybehenoyl)2,3-diacetoxyglycerol.

In one embodiment, the poplar propolis, propolis resin or propolis resin extract has a concentration of at least about 0.1 mg/g, about 0.5 mg/g, about 1 mg/g, about 1.5 mg/g, about 2 mg/g, about 2.5 mg/g, about 3 mg/g, about 3.5 mg/g, about 4 mg/g, about 4.5 mg/g, about 5 mg/g, about 5.5 mg/g, about 6 mg/g, about 7.5 mg/g, about 10 mg/g, about 15 mg/g, about 20 mg/g , about 25 mg/g, about 30 mg/g, about 40 mg/g, about 50 mg/g, about 75 mg/g, about 100 mg/g, about 125 mg/g, about 150 mg/g, about 175 mg/g, about 200 mg/g, 250 mg/g, about 300 mg/g, about 350 mg/g, about 400 mg/g, about 450 mg/g, about 500 mg/g, about 550 mg/g, or about 600 mg/g of methyl 3,8-dihydroxydocosanoate.

In various embodiments, the poplar propolis, propolis resin or propolis resin extract further comprises any combination of two or more of CAPE, chrysin, galangin, pinocytin, pinocytin alcohol, benzylcaffeic acid, benzylferulic acid, benzylisoferulic acid, cinnamic caffeic acid, cinnamic ferulic acid, pinocytin chalcone and caffeic acid.

In various embodiments, the composition comprises at least one other therapeutic agent or is administered separately, simultaneously or consecutively with at least one other therapeutic agent, wherein the at least one other therapeutic agent is preferably an anti-tumor agent, and the anti-tumor agent is preferably selected from anti-tumor food factors, chemotherapeutic agents or immunotherapeutic drugs.

In various embodiments, the skin cancer treatment, therapeutic agent, or anti-tumor agent is effective to induce apoptosis, such as inducing apoptosis in one or more skin cancer cells or one or more tumor cells.

In one embodiment, the composition is a consumable.

In one embodiment, the composition is a composition for topical administration.

In one embodiment, the topical composition includes one or more penetrants, photoprotectants, UV protectants, vitamins, moisturizers, oils, hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fragrances, fillers, pigments, deodorants, or dyes.

In one embodiment, the composition is a medicament or drug.

In various embodiments, the composition is formulated for oral, topical, or parenteral administration.

In one embodiment, a composition formulated for oral administration includes gamma-cyclodextrin.

In one embodiment, the composition formulated for topical administration includes alpha-cyclodextrin.

In one embodiment, the composition formulated for topical administration includes β-cyclodextrin.

In one embodiment, the composition includes one or more additional anti-epithelial cancer drugs.

In one embodiment, the composition is a pharmaceutical composition.

In various embodiments, the chemotherapeutic agent is selected from the group consisting of: mitotic inhibitors, such as vinca alkaloids, including vincristine, vinblastine, vinorelbine, vindesine, vinflunine, podophyllotoxin, paclitaxel (including docetaxel, lalotaxel, ortataxel, paclitaxel and tesetaxel), and epothilones, such as ixabepilone; topoisomerase I inhibitors, such as topotecan, irinotecan, camptothecin, rubitecan and belotecan, topoisomerase II inhibitors, including amsacrine, etoposide, etoposide phosphate and teniposide, anthracyclines, such as aclarubicin, daunorubicin, doxorubicin, epirubicin, flavin, amrubicin, pirarubicin, valrubicin and daurubicin, and anthracenediones, such as mitoxantrone and pixantrone; antimetabolites, including dihydrofolate reductase inhibitors antitumor agents, such as aminopterin, methotrexate, pemetrexed, thymidylate synthase inhibitors, such as raltitrexed and pemetrexed, adenosine deaminase inhibitors, including pentosidine, halogenated or ribonucleotide reductase inhibitors, such as cladribine, clofarabine and fludarabine, thiopurines, including thioguanine and mercaptopurine, thymidylate synthase inhibitors, including fluorouracil, capecitabine, tegafur, camofer and floxuridine, DNA polymerase inhibitors, such as cytarabine, ribonucleotide reductase inhibitors, such as gemcitabine, hypomethylating agents, including azacitidine and decitabine, and ribonucleotide reductase inhibitors, such as hydroxyurea; cell cycle non-specific antitumor agents, including alkylating agents, such as nitrogen mustards, including dichloromethyl diethylamide, cyclophosphamide, ifosfamide, chlorambucil, phenylalanine, chlorambucil Nimustine, bendamustine, uracil mustard, estramustine, nitrosoureas including carmustine, lomustine, semustine, fotemustine, nimustine, ranimustine, and streptozotocin, alkyl sulfonates including busulfan, mannosulfan, and trooxsulfan, aziridines including carboquinone, thiotepa, triazinon, and trotamide, alkylating agents including platinum agents such as cisplatin, carboplatin, oxaliplatin, nedaplatin, 4-nitrotoxin, and oxaliplatin. acid trinuclear platinum, seraplatin, hydrazines such as procarbazine, triazenes such as dacarbazine, temozolomide, hexamethylmelamine, and dibromomannitol, and streptomycins such as actinomycin, bleomycin, daunomycin, mitomycin, and plicamycin; photosensitizers including aminolevulinic acid, methylaminolevulinic acid, ethoxyproline, and porphyrin derivatives such as porfimer sodium, talaporfin, temoporfin, and verteporfin; enzyme inhibitors chemotherapeutic agents, including farnesyl transferase inhibitors such as tipifarnib, cyclin-dependent kinase inhibitors such as alvocidib and seliciclib, proteasome inhibitors such as bortezomib, phosphodiesterase inhibitors such as anagrelide, IMP dehydrogenase inhibitors such as thiazofurine, lipoxygenase inhibitors such as masoprocol, and PARP inhibitors such as olaparib; receptor antagonists such as endothelin receptor antagonists including atrasentan, retinoid X receptor antagonists such as bexarotene and testosterone; and other chemotherapeutic agents including amsacrine, trabectedin, retinoids such as alitretinoin and tretinoin, arsenic trioxide, asparagine depleting agents such as asparaginase or pegaspargase, celecoxib, demeclofenac, ilisimol, elsamitrucin, etoglucagon, and lonidamine.

Reference to a series of numbers (e.g., 1 to 10) disclosed herein is intended to include reference to all rational numbers within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10) and any range of rational numbers within that range (e.g., 2 to 8, 1.5 to 5.5, and 3.1 to 4.7); thus, all subranges of all ranges explicitly disclosed in the present invention are also explicitly disclosed. These are merely examples that are explicitly intended, and all possible combinations of numerical values between the minimum and maximum values listed are to be considered to be expressly stated in this application in a similar manner.

Some patent specifications, other external documents or other information sources are also cited in this specification, mainly as background references when discussing the features of the present invention. Unless otherwise specifically stated, the reference to these external documents shall not be understood as an admission that these documents or information sources are prior art or part of the common general knowledge in the field in any jurisdiction.

In a broad sense, the present invention may also be considered to exist in the parts, elements and features mentioned or indicated in the present specification, either individually or as a whole, and in any or all combinations of two or more of the parts, elements and features, and if specific integers mentioned in the present invention have known equivalents in the art to which the present invention relates, then these known equivalents are incorporated into the present invention as if they were individually set forth.

DETAILED DESCRIPTION

The present invention relates to fatty acid compounds and compositions comprising these dihydroxy fatty acid glycerides having anti-epithelial cancer activity. Pharmaceutical compositions, such as anti-epithelial cancer compositions, have and/or can enhance the efficacy of anti-epithelial cancer, and in certain embodiments can enhance the activity and physicochemical properties of these compositions comprising these compounds, such as propolis and poplar extracts. Nutritional compositions, such as skin and intestinal health compositions have health benefits. In certain embodiments, the activity and physicochemical properties of these compositions are enhanced, such as in the presence of propolis or propolis extracts or components or poplar extracts or components.

In one aspect, the present invention relates to a method for treating or preventing epithelial cancer in an individual, the method comprising administering to an individual in need thereof an effective amount of a composition comprising a therapeutically effective amount of a compound of formula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein R ₂ , R ₃ , R ₄ and R ₅ are independently H or acetyl (CH ₃ CO-),

_R6 is H or _CH3 , _R7 is _CH3 or a C2-C6 saturated or unsaturated hydrocarbon, and

x and y are independently integers ranging from 3 to 14,

Provided that when R ₆ is H, x+y is greater than 10 and less than or equal to 18, and when R ₆ is CH ₃ , x+y is greater than 9 and less than or equal to 17.

In one embodiment, x is greater than or equal to 4. In one embodiment, the present invention relates to a method for treating or preventing epithelial cancer in an individual, the method comprising administering to an individual in need thereof an effective amount of a composition comprising a therapeutically effective amount of at least one compound selected from any one or more of the following groups,

a) 3,8-dihydroxyeicosanoic acid,

b) 1-(3,8-dihydroxyeicosanoyl)propylene glycol

c) 1-(3,8-dihydroxyeicosanoyl)-2-acetoxyglycerol,

d) 1-(3,8-dihydroxyeicosanoyl)3-acetoxyglycerol,

e) 1-(3,8-dihydroxyeicosanoyl)2,3-diacetoxyglycerol,

f) 1-(3,8-diacetoxyeicosanoyl)2,3-diacetoxyglycerol,

g) methyl 3,8-dihydroxyeicosanoate,

h) 3,8-dihydroxyheneicosanoic acid,

i) 1-(3,8-dihydroxyheneicosanoyl)propylene glycol

j) 1-(3,8-dihydroxyheneicosanoyl)2-acetoxyglycerol,

k) 1-(3,8-dihydroxyheneicosanoyl)3-acetoxyglycerol,

1) 1-(3,8-dihydroxyheneicosanoyl) 2,3-diacetoxyglycerol,

m) 1-(3,8-diacetoxyheneicosanoyl)2,3-diacetoxyglycerol,

n) 3,8-dihydroxyheneicosanoic acid methyl ester,

o) 3,8-dihydroxydocosanoic acid,

p) 1-(3,8-dihydroxybehenanoyl)propylene glycol

q) 1-(3,8-dihydroxydocosanoyl)2-acetoxyglycerol,

r) 1-(3,8-dihydroxybehenanoyl)3-acetoxyglycerol,

s) 1-(3,8-dihydroxydocosanoyl)2,3-diacetoxyglycerol,

t) 1-(3,8-diacetoxybehenanoyl)2,3-diacetoxyglycerol

u) 3,8-dihydroxydocosanoic acid methyl ester,

An acceptable salt or solvate.

The compounds and pharmaceutical compositions, e.g., anti-cancer compositions, are used in some embodiments to treat or prevent epithelial cancer, or in other embodiments, e.g., in the presence of propolis, propolis extracts or fractions, propolis resins or propolis resin extracts, or poplar extracts or fractions, to improve skin health, and/or enhance the activity and physicochemical properties of propolis, propolis extracts or fractions, propolis resins or propolis resin extracts, or materials containing propolis.

Therefore, as long as the anti-epithelial cancer compositions are formulated to be suitable for administration to mammalian subjects, for example, these compositions are composed of materials that are safe for humans, they can be used to prepare anti-epithelial cancer pharmaceutical compositions and drugs.

In addition, since the anti-epithelial cancer activity of the embodiments of the composition of the present invention can be maintained for a period of time, the dosage or frequency of administration of the composition can be reduced, or a better therapeutic effect can be produced, or both.

The phrases "anti-epithelial cancer composition" or "composition having anti-epithelial cancer activity" (used interchangeably herein) of the present invention contemplate any type of composition. Examples include anti-epithelial cancer compositions consisting of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and cyclodextrin, or anti-epithelial cancer compositions containing materials such as propolis, propolis resin or propolis resin extracts and cyclodextrin. In particular, synergistic compositions include compositions that enhance any anti-epithelial cancer activity (only observed in propolis or cyclodextrin). In certain embodiments, the anti-epithelial cancer composition may be an anti-basal cell carcinoma, anti-squamous cell carcinoma or anti-melanoma composition. In other embodiments, the anti-epithelial cancer composition is an anti-gastrointestinal cancer composition, such as an anti-colorectal cancer, anti-gastric cancer or anti-laryngeal cancer composition.

The term "and/or" can mean "and" or "or".

The terms "cancer" and "cancerous" refer to the physiological condition of mammals that is typically characterized by abnormal or unregulated cell proliferation, cell survival, cell motility, neoplasia and/or tumorigenicity. Cancer and cancer pathology may be associated with, for example, metastasis, interference with the normal function of neighboring cells, abnormal release of cellular agents or other exudates, suppression or aggravation of inflammation or immune response, neoplasia, precancerous lesions, malignancies, invasion of surrounding or distant tissues or organs, such as lymph nodes, etc. Specifically included are basal cell carcinoma, squamous cell carcinoma, melanoma and precancerous conditions, which may include dermal tumors, epithelial tumors, oral tumors, such as oral squamous cell carcinoma, carcinoma in situ, and invasive basal cell carcinoma, squamous cell carcinoma or melanoma cancers, and secondary tumors derived therefrom. Also specifically included are gastrointestinal cancers, such as colorectal cancer and precancerous conditions, which may include epithelial tumors, non-epithelial tumors, cancers, such as carcinoma in situ, and invasive colorectal cancer. Also included are gastric cancers and precancerous conditions, including epithelial tumors, adenocarcinomas, gastric lymphomas, carcinoid tumors, stromal tumors. Also included are laryngeal cancers and precancerous conditions, including epithelial tumors, squamous cell carcinomas, adenocarcinomas. Likewise, mucosal tissue cancers are particularly contemplated. Cancers can be, for example, carcinoma in situ, as well as invasive cancers.

The term "comprising" used in this specification means "consisting at least of a part". When understanding the sentences containing this term in this specification, the features beginning with this term in each sentence need to be present, but other features may also be present. The related verb conjugations of this term are understood in the same way.

An "effective amount" is the amount required to achieve a therapeutic effect. Freireich et al. (1966) described the interrelationship of animal and human doses (based on milligrams per square meter of body surface area). Body surface area can be roughly determined based on the height and weight of an individual. See, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardley, New York, 1970, 537. As will be appreciated by those skilled in the art, effective doses vary depending on route of administration, excipient use, etc.

The poplar "extract" or "component" used in the present invention is suitable for use in the present invention if it at least retains one or more anti-epithelial cancer activities, or includes one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof. Exemplary extracts or components of poplar used in the present invention are rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof. Examples of these functional extracts include the anti-epithelial cancer preparations described in the Examples. It should be understood that the poplar extract or component rich in one or more compounds of formula (I) can be increased or decreased during preparation, for example, at least partially separated or purified compounds of formula (I) can be added to a composition, such as poplar or poplar extract or component, or removed from poplar or poplar extract or some compounds other than one or more compounds of formula (I).

The propolis "extract" or "component" used in the present invention is suitable for the present invention if it at least retains one or more anti-epithelial cancer activities or efficacies possessed by propolis, and/or includes one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof. Exemplary extracts or components of propolis used in the present invention are rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof. Examples of these functional extracts include the anti-epithelial cancer tinctures described in the embodiments. It should be understood that the propolis extract or component rich in one or more compounds of formula (I) can be increased or decreased during preparation, so that at least partially separated or purified compounds of formula (I) can be added to the composition, such as propolis or propolis extracts or components, or removed from the components of propolis or propolis extracts or compounds instead of one or more compounds of formula (I).

Chromatography as used herein refers to a separation process in which compounds dissolved or dispersed in a solvent phase or otherwise transferred are contacted with a solid phase so that the solid phase selectively delays one or more compounds based on the presence of other compounds in the solvent, thereby enabling their separation. Examples of various types of chromatography include column chromatography, liquid chromatography, gas chromatography, supercritical chromatography, size exclusion chromatography, preparative chromatography, solid phase extraction. Chromatography can be used to determine the concentration of one or more compounds in a mixture, identify unknown compounds, and/or separate one or more compounds, such as to obtain detailed structural analysis data, for use as analytical standards and/or for use in biological assays.

Methods and tests for determining one or more biological effects produced by one or more compounds or compositions of the present invention (e.g., including one or more compounds of formula (I), and optionally, compositions including propolis or poplar extracts) are well known in the art, and relevant examples are also described in the present invention; and such methods and tests can be used to identify or verify efficacy, for example, the efficacy of one or more compounds or compositions of the present invention, including compositions containing one or more functional extracts or functional components of propolis, such as propolis, propolis extracts, poplar extracts, or propolis or poplar components rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof. For example, in order to determine the ability of one or more compounds or compositions of the present invention to increase one or more oncogene features in cells, as described in the examples of the present invention, it is necessary to verify the efficacy of one or more compounds or compositions of the present invention, or to identify one or more functional extracts or functional components of propolis or poplar.

"Propolis" used in the present invention includes propolis obtained and produced by bees through any plant source. In one embodiment, the propolis refers to "poplar type" propolis. "Poplar type" propolis also has different names, such as "poplar" propolis. For example, the propolis is mainly derived from the buds and leaf exudates of one or more poplar varieties, followed by birch, larch or willow. Plant sources allow characteristic compounds to exist, and propolis is divided into seven major categories according to plant sources (Sforcin and Bankova, 2011. Propolis: Is there a potential for new drug development? Journal of Ethnopharmacology, 133: 253-260.). These categories are European, North American, southern South American, and New Zealand "poplars," which are extremely high in flavonoid aglycones such as chrysin and galangin; "Brazilian green" poplars, which contain isopentenyl p-coumaric acids such as aspirin C; Russian "birch," also rich in flavonoid aglycones such as apigenin, rhamnetin, and kaempferol; "red propolis," from the Cruci variety of Cuba, Brazil, Mexico, and Venezuela, which contains polyisopentenyl benzophenones, including nemorosone and garcinol; "Mediterranean" poplars from Greece, Sicily, Crete, and Malta, which are rich in diterpenes derived from conifers; and "Pacific" poplars from Okinawa, Taiwan, and Indonesia, which contain "propolis."

Due to the complex and multi-component nature of propolis resin, it is challenging to identify and verify the presence of anti-cancer components in propolis resin. It has been shown that "Poplar" propolis resin contains a variety of compounds, including sugars, glycerol, simple fatty acids, volatile oils and phenolic acid glycerides. The present invention describes a group of compounds in propolis that have not been previously reported and whose anti-cancer activity is hitherto unknown. These compounds are long-chain fatty acids and their equivalent glycerides. In addition, these dihydroxy free fatty acids and glycerides can be partially acetylated, up to fully acetylated. These compounds have the structure of formula (I):

wherein R ₂ , R ₃ , R ₄ and R ₅ are independently H or acetyl (CH ₃ CO-),

_R6 is H or _CH3 , _R7 is _CH3 or a C2-C6 saturated or unsaturated hydrocarbon, and

x and y are independently integers of 3 to 14,

Provided that when R ₆ is H, x+y is greater than 10 and less than or equal to 18, and when R ₆ is CH ₃ , x+y is greater than 9 and less than or equal to 17.

It will be appreciated by those skilled in the art that the characteristics of propolis and, in some cases, suitability for specific uses, including uses in the present invention, can be determined by analyzing the components of propolis. The presence and quantity of specific compounds (including compounds discussed in the present invention) - that is, "marker compounds" - can determine whether a specific propolis source, such as poplar propolis, is suitable for a specific use compared to Brazilian propolis. In certain embodiments of the present invention, before preparing the composition, as a preliminary classification step, the presence or content of one or more marker compounds can be determined or analyzed. In other embodiments, the presence or content of one or more marker compounds is determined and reported, such as in an indication related to the composition of the present invention, such as an efficacy or activity indication.

"Instructions" may take any form, including, but not limited to, a label that is part of the product, such as a label etched or printed on the surface of the product or composition, instructions printed or etched on a capsule or tablet, packaging or labeling associated with the product, such as a label affixed to or incorporated into a package containing the product or composition, information accompanying the product or composition but separate from the product or composition, such as a certificate of analysis; a website, brochure, brochure or presentation material related to the sale or marketing of the product or combination.

Can use method known in the art to separate and/or purify formula (I) compound from poplar type propolis.The present invention provides exemplary methods.Similarly, can also use method known in the art to separate and/or purify formula (I) compound from plant source such as poplar and poplar extract or exudate.The present invention also provides exemplary methods.These methods are also applicable to determine the concentration or amount of one or more formula (I) compounds or its pharmaceutically acceptable salt or solvate present in the composition of the present invention, for example, in order to report the concentration or amount.

In addition, the compound of formula (I) can be prepared by synthesis using methods known in the art. For example, when synthesizing the compound of formula (I), it is particularly contemplated to use known raw materials and synthesize by Grignard reaction (Smith, Michael B.; March, Jerry (2007), "Advanced Organic Chemistry: Reactions, Mechanisms and Structures" (6th Edition), New York: Wiley-Interscience Press, ISBN 0-471-72091-7). For example, it is contemplated to prepare the compound of formula (I) using raw material 3,8-dihydroxyoctanoic acid and a Grignard reagent selected from dodecylmagnesium bromide, tridecylmagnesium bromide or tetradecylmagnesium bromide. These Grignard reagents can be prepared with the corresponding long-chain fatty alcohol dodecan-1-ol (lauryl alcohol), 1-tridecanol or 1-tetradecyl alcohol (myristyl alcohol). It is also contemplated to selectively oxidize the primary alcohol of 3,8-dihydroxyoctanoic acid to aldehydes to achieve Grignard coupling of the raw materials. The resulting 3,8-dihydroxy fatty acid can then be esterified using a suitable lipase or by standard chemical esterification with glycerol, glycerol-2-acetate or glycerol 2,3-acetate to give the main compound of formula (I).

When used in connection with the compositions of the invention or components of such compositions, the phrases "enhanced activity" or "enhanced anti-epithelial cancer activity" and grammatical synonyms and derivatives thereof are intended to indicate that, when present in the composition, an equivalent amount or equivalent concentration of the anti-epithelial cancer agent has enhanced anti-epithelial cancer activity compared to the agent not present in the composition (e.g., an isolated agent), and/or that the stability of the composition is improved relative to the individual components and/or that the bioavailability of the composition is improved relative to the individual components. For example, the enhanced activity is at least about 105%, 110%, 115%, 120%, 125%, 130%, 135%, 140%, 145%, 150%, 155%, 160%, 165%, 170%, 175%, 180%, 185%, 190%, 195%, 200% or more of the original activity, and an effective range can be selected between any of these values (e.g., about 105% to about 200%, about 120% to about 200%, about 140% to about 200%, about 150% to about 200%, about 180% to about 200%, and about 190% to about 200%). In certain embodiments, the composition may have enhanced anti-epithelial cancer activity, i.e., at least about 105%, 110%, 115%, 120%, 125%, 130%, 135%, 140%, 145%, 150%, 155%, 160%, 165%, 170%, 175%, 180%, 185%, 190%, 195%, 200% or more of the anti-epithelial cancer activity of propolis alone or cyclodextrin alone. Similarly, by using the method of the present invention, the preferred composition can maintain enhanced anti-epithelial cancer activity, which can also be understood as maintaining enhanced anti-epithelial cancer activity. Without being bound by theory, the enhanced activity (including maintaining enhanced activity) is due to the synergistic effect between the various components of the composition, or due to, for example, improved water dispersibility, making the composition more stable or biopharmaceutical efficiency increased.

The term "oral" includes buccal administration, enteral administration and intragastric administration.

The term "parenteral administration" includes, but is not limited to, topical administration (including administration through any skin, epithelial or mucosal surface), subcutaneous administration, intravenous administration, intraperitoneal administration, intramuscular administration, and intratumoral administration (including any direct administration to a tumor).

The term "pharmaceutically acceptable carrier" is intended to refer to a carrier, including but not limited to an excipient, diluent or adjuvant that can be administered to an individual as a component of the composition. Preferably, the carrier does not reduce the activity of the composition and is not toxic when administered in a dose sufficient to release an effective amount of propolis or an extract thereof, or when administered with another anti-epithelial cancer drug.

The "s" after a noun refers to the singular or plural form, or both.

The term "subject" is intended to refer to an animal, preferably a mammal, more preferably a mammalian companion animal or a human. Preferred companion animals include cats, dogs and horses. Other mammalian subjects refer to agricultural animals, including horses, pigs, sheep, goats, cattle, deer or poultry, or laboratory animals, including monkeys, rats or mice.

The term "treat" and its derivatives should be understood in the broadest possible context. The term should not be construed to imply that an individual is treated until full recovery. Thus, "treat" broadly includes maintaining an individual's disease progression or symptoms at a substantially static level, increasing an individual's rate of recovery, preventing the onset of symptoms and/or ameliorating the severity of a particular condition, or prolonging the patient's life. The term "treat" broadly also includes maintaining the physical health of susceptible individuals and enhancing resistance to disease.

Exemplary Uses of the Invention

The methods and compositions described herein are useful for treating or preventing epithelial cancers, neoplastic diseases associated with epithelial cells, and symptoms of such cancers, including symptoms of cancer treatment and related diseases.

In one example, the methods and compositions described herein can be used to treat or prevent skin cancer, such as melanoma, tumor diseases associated with melanoma, and symptoms of melanoma, melanoma treatment, and related diseases. Melanoma (also known as malignant melanoma) is a tumor disease that affects melanocytes.

Melanoma is the least common, but most aggressive and threatening form of skin cancer. When possible, the preferred treatment is total surgical removal, which is curative if metastases have not occurred.

The methods and compositions described herein can be used to treat or prevent basal cell carcinoma, tumor diseases associated with basal cell carcinoma, and symptoms of basal cell carcinoma, basal cell carcinoma treatment, and related diseases. Basal cell carcinoma is a tumor disease that affects the basal cells of the dermis.

Basal cell carcinoma develops in the lowest layer of the epidermis. When possible, the preferred treatment is total removal through surgery, which is curative.

In certain embodiments, the methods and compositions are used to treat or prevent squamous cell carcinoma, neoplastic diseases associated with squamous cell carcinoma, and symptoms of squamous cell carcinoma, squamous cell carcinoma treatment, and related diseases.

Squamous cell carcinoma is a tumor that develops in the cells in the middle layer of the epidermis. Although less common than basal cell carcinoma, it is more likely to metastasize and can be fatal if untreated.

The present invention provides methods and compositions for inhibiting skin tumor formation, skin tumor growth, skin tumor metastasis, or treating or preventing skin cancer in a desired individual. Without being bound by theory, applicants believe that at least partial inhibition may be achieved, for example, by preventing UV-induced DNA damage or preventing or reducing the formation of reactive oxygen species.

In certain embodiments, the present invention also relates to a method of at least partially reversing resistance of tumor cells in an individual suffering from skin cancer to skin cancer treatment, or to a method of fully or partially reversing resistance of a patient suffering from skin cancer to skin cancer treatment, a method of resensitizing one or more tumors in an epithelial cancer patient, wherein the one or more tumors are resistant to epithelial cancer treatment, or are expected to be resistant to epithelial cancer treatment, the method comprising the step of administering to the patient a composition comprising, or consisting essentially of, or consisting of propolis, propolis resin or propolis resin extract enriched in one or more compounds of formula (I) and cyclodextrin.

In one embodiment, the one or more tumors are resistant to skin cancer treatment, are predicted to be resistant, or are likely to become resistant due to increased activity of one or more procancerous cell survival signaling pathways in one or more tumors or in the patient, including increased activity in one or more AKT, JNK, or JAK/STAT signaling pathways, for example, increased activity in a sample from the patient (e.g., a tissue sample, tumor biopsy, or a blood or plasma sample).

Precancerous cell survival signaling pathways associated with the development and progression of skin cancer are known in the art.

The methods and compositions are useful for treating or preventing melanoma, neoplastic diseases associated with melanoma cells, and symptoms of melanoma, melanoma treatment, and related diseases.

The most common treatments are surgery to remove the tumor and radiation therapy. Chemotherapy drugs may be used in combination with surgery and/or radiation therapy.

The methods and compositions are also useful for maintaining or improving skin health.

This includes treating or preventing diseases associated with poor skin health, low immunity, and skin inflammation. For example, the methods and compositions can be used to treat or prevent skin aging, sunburn, dermatitis, eczema, psoriasis, ichthyosis, and related inflammatory diseases, as well as treating or preventing skin redness, inflammation, dryness, cracking, or itching.

Propolis and Propolis Materials

Propolis is available in New Zealand and elsewhere, usually as a resinous, viscous solid. Propolis can be obtained from beehives and stored in warehouses, for example, for assessing the propolis content. Propolis or its extracts are generally processed into fine granules or concentrated tinctures. Various methods for preparing active propolis or its extracts into granules or concentrated tinctures are known. Most commonly, natural propolis is extracted with ethanol or an ethanol/water mixture to make a dilute tincture. The wax associated with natural propolis is the most difficult to dissolve in solvents, so it is essentially not extracted. Any extracted wax can be removed by cooling the dilute tincture and then settling, filtering or centrifuging. The solvent can then be completely evaporated by partially concentrating the tincture to obtain a concentrated tincture, optionally followed by freeze drying to obtain a powder. Or the tincture can be spray-dried to obtain a powder. The components can be prepared using methods known in the art, such as chromatography (such as HPLC) using a size exclusion matrix or a reversed phase matrix, or supercritical separation. The typical solvent used for this chromatography is ethanol or another alcohol that is miscible with water.

In one embodiment, propolis or concentrated propolis tincture is combined with other compounds that enhance the properties of propolis, such as compounds that enhance preparation or administration convenience, or compounds that enhance anti-epithelial cancer activity, or compounds that enhance the stability of one or more anti-epithelial cancer activities present in propolis. The examples of other compounds are those compounds that improve the therapeutic effect of propolis. Specifically considered as an exemplary composition, one or more compounds that are only found in propolis, particularly poplar-type propolis, are added, including bioactive compounds such as CAPE, caffeic acid, pinocytogenin, benzyl caffeic acid, cinnamic caffeic acid, benzyl ferulic acid, cinnamic ferulic acid, chrysin, galangin, pinocytogenin, pinocytogenin chalcone and pinocytogenin-3-acetate. In other examples, other compounds that improve or maintain the physiological effects of the composition are included, such as mannitol can be added to enhance the diuretic effect of the resulting composition. Or other compounds, such as excipients and/or propellants, can be added to improve the dosage, manufacturability or delivery of the composition.

In particularly contemplated embodiments, dewaxed propolis resin, optionally with one or more other compounds, is included with cyclodextrin, and the mixture is dried. The mixture is further processed, for example, to obtain a particle size distribution that allows for mixing with other components of the composition, for ease of tableting, or for ease of administration to an individual.

In typical embodiments, propolis or propolis resin is sterilized, for example by heating to kill bacteria, protozoa, yeasts, fungi and other organisms naturally present in the propolis.

Poplar Extract

As described in the examples of the present invention, poplar (Populus) extracts, especially exemplary compounds of formula (I) in bud, leaf and twig exudates, have been identified. Without being bound by theory, the applicant believes that the compounds of the present invention are present in poplar-type propolis and propolis resins or are associated with poplar-type propolis and propolis resins. It will be appreciated by those skilled in the art that, for use in the present invention, poplar or propolis extracts may be processed into a form suitable for further processing and encapsulation, such as containing cyclodextrin, or extracting poplar or propolis extracts with solvents (e.g., alcohol) while maintaining biologically active ingredients. Poplar or propolis extracts are generally processed into fine granules or concentrated tinctures.

Cyclodextrin and cyclodextrin materials

Cyclodextrins are cyclic molecules composed of glucopyranose ring units forming a ring structure. Cyclodextrin molecules are water soluble due to the hydrophobic interior and hydrophilic exterior of the molecule. The solubility of a cyclodextrin can be altered by substitution of the external hydroxyl groups. Likewise, the hydrophobicity of the interior can be altered by substitution, although the hydrophobicity of the interior generally allows for the accommodation of relatively hydrophobic guests within the cavity. The presence of one molecule within another is called complexation, and the resulting product is called an inclusion complex. Cyclodextrins are generally identified by reference to the number of monomeric units that make up the molecule, with α-cyclodextrin comprising six monomeric units, β-cyclodextrin comprising seven monomeric units, and γ-cyclodextrin comprising eight monomeric units. Larger cyclodextrin molecules have been described, including well-characterized cyclodextrins containing 32 1,4-anhydroglucopyranose units.

The cyclodextrin molecule can be conveniently derivatized, for example by modification, to alter one or more of its physicochemical properties. Examples of cyclodextrin derivatives include methylated cyclodextrins, sulfobutyl cyclodextrins, maltosyl cyclodextrins, hydroxypropyl cyclodextrins, such as β-hydroxypropyl cyclodextrin and γ-hydroxypropyl cyclodextrin, and salts thereof. It will be appreciated by those skilled in the art that various derivatives of cyclodextrins may be suitable for specific purposes, for example, certain derivatives of cyclodextrins may not be administered to humans but are suitable for industrial use.

The cyclodextrins comprising the anti-epithelial cancer composition of the present invention are commercially available or can be prepared separately by methods well known to those skilled in the art. It is obvious to those skilled in the art that the cyclodextrins in the anti-epithelial cancer composition for individual administration, such as the cyclodextrins used to make beverages, foods or medicines, should be safe for the human body, and preferably pharmaceutically acceptable cyclodextrins.

In particularly contemplated embodiments, alpha-cyclodextrin, gamma-cyclodextrin or a combination comprising alpha-cyclodextrin, gamma-cyclodextrin or both is used. In these embodiments, as described in the examples of the present invention, anti-epithelial cancer activity is significantly enhanced. These compositions comprising alpha-cyclodextrin and/or gamma-cyclodextrin can be formulated to have a better mouthfeel or palatability, for example, a composition comprising alpha-cyclodextrin and/or gamma-cyclodextrin and propolis covers any unpleasant taste in propolis.

Cyclodextrins suitable for use in the present invention may be obtained from commercial sources or prepared separately by methods known in the art, such as from starch by enzymatic conversion. In certain embodiments, CAVAMAX W6, W7 or W8 foods, alpha, beta or gamma cyclodextrins sold by Wacker AG, are used.

In certain embodiments, such as those directed to topical administration of the composition, non-food grade cyclodextrins are used, for example, substituted cyclodextrins including those within the CAVASOL range are contemplated.

Epithelial cancer

Epithelial cancers suitable for treatment with the composition of the present invention include squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, large cell carcinoma, small cell carcinoma and transitional cell carcinoma. It should be understood that cancer is a term commonly used for malignant epithelial tumors. There are two main types of cancer, classified by the type of epithelium from which they originate. These are squamous cell carcinomas that originate from squamous epithelium and adenocarcinomas that originate from glandular epithelium.

In various embodiments, the epithelial cancer is selected from gastric cancer (intestinal type), gastric cancer (diffuse/mucinous), moderately differentiated colon cancer, poorly differentiated colon cancer (mucinous), hepatocellular carcinoma (including poorly differentiated hepatocellular carcinoma), renal cell carcinoma (Gravetz tumor), endometrioid carcinoma, breast cancer (including invasive breast cancer), metastatic cancer (lymph node), colorectal cancer, oral cancer (including esophageal cancer, pharyngeal cancer or laryngeal cancer), skin cancer (including basal cell skin cancer, squamous cell skin cancer and melanoma) and ovarian cancer.

It is specifically contemplated that the methods and compositions described herein are used to treat oral or laryngeal cancer, also known as esophageal cancer, pharyngeal cancer, or laryngeal cancer, including tumors arising in the tissues of the pharynx, nasopharynx, oropharynx, hypopharynx, larynx (larynx), or tonsils. Likewise, treatment of gastric cancer, as well as basal cell skin cancer, squamous cell skin cancer, and melanoma are specifically contemplated.

Composition

Exemplary anti-epithelial cancer compositions include one or more compounds of Formula (I) or one or more pharmaceutically acceptable salts or solvates thereof and a pharmaceutically acceptable carrier.

Compositions suitable for individual administration can be formulated into medical products, medical devices, medicaments or drugs. Those skilled in the art can prepare suitable formulations with reference to the techniques and teachings of this specification.

The compositions useful in the present invention can be formulated for administration to an individual by a selected route, including but not limited to oral or parenteral (including topical, subcutaneous, intramuscular and intravenous). It will be appreciated by those skilled in the art that the route of administration to an individual will generally take into account the purpose for which the composition is administered - for example, to improve skin health or to treat or prevent skin cancer, the route of administration of the pharmaceutical composition will generally take into account health factors or the nature of the skin cancer.

Typically, when taken orally, the pharmaceutical compositions useful in the present invention can be formulated by those skilled in the art according to known formulation techniques. In certain embodiments, the composition formulated for oral administration comprises γ-cyclodextrin. In certain embodiments, the composition formulated for oral administration comprises α-cyclodextrin.

Thus, pharmaceutical compositions useful according to the invention can be formulated with suitable pharmaceutically acceptable carriers (including excipients, diluents, adjuvants and combinations thereof) selected according to the likely route of administration and standard pharmaceutical practice. See, for example, Remington's Pharmaceutical Sciences, 16th edition, Osol, A. ed. Mack Publishing Co., Ltd., 1980.

Although the suitable route of administration of certain embodiments, for example, the route of administration including one or more compounds of formula (I) is oral, it should be understood that for any composition, any route of administration is suitable, including administration by multiple routes, and the routes of administration of different agents are different. Therefore, nasal mucosal administration (nasal or buccal inhalation) and vaginal and rectal administration of any composition are also considered. Intramedullary, epidural, intra-articular and intrapleural administration of any composition are also considered. Compositions administered by a first route of administration and a second route of administration are also considered, optionally, including at least one other anti-epithelial cancer factor, wherein the first route of administration is suitable for administering one or more other agents alone, simultaneously or continuously, including one or more other anti-epithelial cancer agents; the second route of administration is an oral composition, and at least one other anti-epithelial cancer agent is administered topically at the same time.

The composition can also be formulated into dosage forms. The dosage forms of the present invention can be taken orally in the form of powders, liquids, tablets or capsules. Suitable dosage forms can contain additives as needed, including emulsifiers, antioxidants, flavorings or coloring agents, or have enteric coatings. Suitable enteric coatings are known. Enteric coatings coat the active ingredient and prevent the active ingredient from being released in the stomach, but allow the dosage form to be released after leaving the stomach. Useful dosage forms in the present invention can be suitable for quick release, sustained release, regulated release, sustained release, pulsed or controlled release of the active ingredient. Suitable preparations can contain additives as needed, including emulsifiers, antioxidants, flavorings or coloring agents.

Capsules may contain any standard pharmaceutically acceptable material, such as gelatin or cellulose. Tablets may be prepared by compressing a mixture of the active ingredient with a solid carrier and a lubricant according to conventional procedures. Examples of solid carriers include starch and sugar bentonite. The active ingredient may also be administered in the form of a hard-shell tablet or capsule containing a binder such as lactose or mannitol, conventional fillers and compressed tablets. The pharmaceutical composition may also be administered parenterally. Examples of injectable pharmaceutical forms include aqueous solutions of the active agent, isotonic saline or 5% glucose, or other well-known pharmaceutically acceptable excipients. Solubilizers well known to those skilled in the art may be used as pharmaceutical excipients for releasing anti-epithelial cancer drugs.

Injectable dosage forms can be formulated as liquid solutions or suspensions. Solid forms suitable for dissolving in liquids or suspending in liquids prior to injection can also be prepared. The dosage form can also be emulsified. The one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof can be mixed with a carrier, such as water, saline, glucose, glycerol, ethanol, etc. and combinations thereof.

Sustained-release preparations can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers, the matrix containing one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and optionally, containing propolis and/or poplar extract and/or cyclodextrin, and also containing at least one other anti-epithelial cancer drug (if any). The matrix can be in the form of a shaped article, such as a film or a microcapsule. Examples of sustained-release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactides (see US 3,773,919), copolymers of L-glutamic acid and ethyl L-glutamate, non-degradable ethylene-vinyl acetate and degradable lactic acid-glycolic acid copolymers, such as LUPRON DEPOT ^™ (injectable microspheres composed of lactic acid-glycolic acid copolymers and leuprolide acetate).

Particularly contemplated are topical preparations comprising compositions, such as preparations comprising one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or concentrated propolis, propolis resin or propolis resin extract and cyclodextrin, and/or preparations when at least one other anti-epithelial cancer drug is present. Topical preparations can be prepared as lotions, creams, ointments, pastes or salves using known carriers. In certain embodiments, compositions formulated for topical administration include α, β or γ-cyclodextrin.

In certain embodiments, the topical formulation includes one or more penetrants, such as one or more alkyl lactates, one or more antioxidants, such as vitamin E (α-tocopherol) or another naturally occurring antioxidant, including polyphenolic antioxidants, such as proanthocyanidins and chlorogenic acid, quinic acid, and ferulic acid, one or more photoprotectants or UV protectants, such as titanium dioxide or carnosic acid, one or more lipids, collagen, keratin, or other proteins.

In certain embodiments, the topical composition includes one or more common carriers in cosmetics, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fragrances, fillers, UV protectants, pigments, deodorants and dyes. Typically, the composition will contain the amount conventionally used in the art, such as 0.01% to 20% relative to the total weight of the composition. These carriers are added to the lipid phase, aqueous phase or one or more phases, vesicles, such as one or more lipid vesicles, microparticles or other components of the topical preparation according to the properties and specific embodiments.

In certain embodiments, particularly when the composition is an emulsion, the lipid/fat phase may account for 5% to 80% (by weight), for example 5% to 50% (by weight) of the total weight of the composition. The oil, emulsifier and co-emulsifier that can be used in the composition in the form of an emulsion are selected from the oils, emulsifiers and co-emulsifiers conventionally used in the art. When emulsifiers and co-emulsifiers are present in the composition, they account for 0.3% to 30% (by weight), for example 0.5 to 20% (by weight) of the total weight of the composition.

In certain embodiments, the composition includes one or more oils, such as one or more mineral oils (liquid petrolatum), vegetable oils (avocado oil or soybean oil), animal oils (lanolin), synthetic oils (perhydrosqualene), silicone oils (cyclomethicone) and fluoro oils, including perfluoropolyethers. Fatty alcohols, such as cetyl alcohol, fatty acids and waxes (carnauba wax or ozokerite) are also used as fatty substances in certain embodiments.

In certain embodiments, the emulsifier and co-emulsifier are fatty acid esters of polyethylene glycol (eg, PEG stearate) or fatty acid esters of glycerol (eg, glyceryl stearate) or a mixture of the two.

Hydrophilic gelling agents are contemplated for use in certain formulations, where such agents include carboxyvinyl polymers (e.g., carbomers), acrylic acid copolymers (e.g., acrylates/alkyl acrylates copolymers), polyacrylamides, polysaccharides, natural gums and clays, as well as lipophilic gelling agents (including modified clays such as bentonite), fatty acid metal salts, hydrophobic silica, and polyethylene.

In certain embodiments, moisturizing creams Dermabase, Unibase, and Vanicream are representative examples of commercially available base creams used as pharmaceutically acceptable carriers.

In certain embodiments, the topical formulation further contains a moisturizer, a depigmenting or pigmenting agent, an antimicrobial agent, or a free radical scavenger.

In certain embodiments, the topical composition is formulated as an aqueous preparation, such as a water-soluble skin cream (water-in-oil or oil-in-water emulsion).

Such formulations may be applied directly, such as by applying directly to a wound, spraying onto a surgical site, etc., or may be applied indirectly, such as by dipping into a bandage or dressing or spraying onto surgical equipment, dressings, and the like.

Also provided are parenteral unit dosage forms comprising one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, optionally with at least one additional therapeutic agent.

In one example, the anti-epithelial cancer composition is a propolis tincture, such as a propolis tincture enriched in one or more compounds of formula (I) or a pharmaceutically acceptable salt or solvate thereof, mixed with cyclodextrin and one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, then adding water and homogenizing the composition, and then spray drying or freeze drying to obtain a powder. Other exemplary anti-epithelial cancer compositions of the present invention include solutions, for example, including the following solutions, wherein the propolis tincture or its components are mixed with cyclodextrin and then dispersed in water; propolis or a material containing propolis and cyclodextrin are dissolved or dispersed in water separately, and then mixed (e.g., kneaded); and first dissolving propolis powder or resin in another organic solvent, or an organic solvent-water mixture that can dissolve propolis powder or resin, such as ethanol, propylene glycol, ethyl acetate, isopropanol, and a mixture thereof with water, and mixing the resulting solution with cyclodextrin, adding water and further mixing (e.g., kneading), and then drying by means known in the art (such as spray drying or freeze drying). Alternatively, water and cyclodextrin may be mixed under conditions where the solubility is equal to or lower than the solubility of cyclodextrin in water, and then the propolis tincture may be mixed therein, and then water may be added and mixed, and then the resulting dispersion may be dried by methods known in the art, such as spraying or freeze drying. In certain embodiments, for example, the above-mentioned anti-epithelial cancer composition prepared as a powder may maintain a stronger anti-epithelial cancer activity, or may maintain the anti-epithelial cancer activity for a longer period of time, compared to the solution of the above-mentioned anti-epithelial cancer composition prepared as a powder, so the above-mentioned anti-epithelial cancer composition prepared as a powder may be preferred.

As long as the expected anti-epithelial cancer activity is achieved, the propolis, propolis resin or propolis resin extract rich in at least one compound of formula (I) of the present invention and cyclodextrin can be in any content. Similarly, as long as the expected anti-epithelial cancer activity is achieved, the compound of formula (I) of the present invention, propolis, propolis resin or propolis resin extract and cyclodextrin can be in any content.

Without being bound by theory, the applicant believes that when the molar ratio of the propolis, propolis resin or propolis resin extract to cyclodextrin is not greater than 1:1, the propolis, propolis resin or propolis resin extract enriched in at least one compound of formula (I) in the composition will be completely included.

In some embodiments, the molar ratio of propolis, propolis resin or propolis resin extract enriched with at least one compound of formula (I) to cyclodextrin in the composition may be greater than 1: 1. In these compositions, excess propolis, propolis resin or propolis resin extract will not be included by cyclodextrin.

Other well-known anti-epithelial cancer substances can be combined with the anti-epithelial cancer composition according to the use of the composition.

Without being bound by theory, the applicant believes that the enhanced anti-epithelial cancer activity observed in the exemplary compositions of the present invention comprising one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof and cyclodextrin is at least partially due to the synergistic effect between the one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, particularly when present as propolis resin or a concentrated fraction of propolis resin and cyclodextrin. In certain embodiments, the exemplary compositions having a synergistic effect are compositions comprising one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof (e.g., a propolis extract or fraction, a propolis resin extract, or a poplar extract or exudate enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof) and α-cyclodextrin. In another contemplated embodiment, the synergistic composition is a composition comprising one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof (e.g., a propolis extract or fraction, a propolis resin extract or a poplar extract or exudate enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof) and γ-cyclodextrin.

In one embodiment, the present invention relates to propolis, propolis resin or propolis resin extract enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and cyclodextrin (e.g., propolis and α-cyclodextrin and/or γ-cyclodextrin), optionally together with at least one anti-epithelial cancer drug for use in the preparation of a medical device, medical product, medicament or medicine.

In one embodiment, the exemplary composition comprising propolis, propolis resin, propolis resin extract, poplar extract or cyclodextrin is formulated for oral administration. In another embodiment, the composition is formulated for parenteral administration, including topical administration. In certain embodiments, the composition is used to induce apoptosis, treat or prevent skin cancer, maintain or improve skin health or one or more other uses mentioned above.

In one embodiment, the composition is a powder, tablet, caplet, pill, hard or soft capsule, or lozenge.

In one embodiment, the composition is a sachet, a dispensable powder, a granule, a suspension, an elixir, a liquid, a beverage, or any other form that can be added to food or a beverage (including water or juice). In one embodiment, the composition is an enteral product, a solid enteral product, or a liquid enteral product.

In one embodiment, the composition may be a cream, ointment, paste, drops (including eye drops or ear drops), an inhalant or inhalable composition, a dressing, a pad or a spray.

In one embodiment, the composition further comprises one or more components (eg, antioxidants) that can prevent or reduce degradation of the composition during storage or after administration.

Particularly contemplated are compositions that additionally comprise milk or one or more milk products or components of milk, such as milk protein, whey protein, colostrum, milk fat, or any component of milk or one or more milk products or components of milk, such as a milk fat component, a milk protein component, a whey protein component, a colostrum component, etc.

Compositions useful in the present invention may also include other factors such as calcium, zinc, magnesium, selenium, vitamin C, vitamin D, vitamin E, vitamin K2, complex carbohydrates, edible oils (including palm oil, olive oil, soybean oil, rapeseed oil, corn oil, sunflower oil, safflower oil, peanut oil, grapeseed oil, sesame oil, walnut oil, almond oil, cashew oil, hazelnut oil, macadamia oil, pecan oil, pistachio seed oil and walnut oil) and other foods (acai, amaranth, apricot, Moroccan nuts, artichoke, avocado, babassu, beans, black currant seeds, borage seeds, Borneo tallow nuts, gourd, buffalo gourd, carob (algaroba), feather palm fruit, coriander seeds, evening primrose, false flax, hemp, kapok seeds, flat-stalked grass, white meadowfoam seeds, mustard, okra seeds (hibiscus seeds), perilla seeds, Brazilian nutwood, pine nuts, poppy seeds, prunes, pumpkin seeds, quinoa, black sesame, rice bran, tea (camellia), thistle, watermelon seeds or wheat germ oil or a combination thereof).

In various embodiments, the composition may include at least one other therapeutic agent, wherein the at least one other therapeutic agent is an antibiotic, such as an aminoglycoside antibiotic (amikacin, gentamicin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin, or paromomycin); ansamycin antibiotics (geldanamycin or herbimycin); carbacephem antibiotics (loracef); carbapenem antibiotics (ertapenem, doripenem, imipenem/cilastatin, or meropenem); first generation cephalosporin antibiotics (cefadroxil, cefazolin, cephalothin, or cephalexin); second generation cephalosporin antibiotics (cefadroxil, cefazolin, cephalothin, or cephalexin); cephalosporin antibiotics (cefixime, cefdinir, cefditoren, cefoperazone, ceftriaxone, cefpodoxime, ceftazidime, ceftibuten, cefuroxime or ceftriaxone); fourth-generation cephalosporin antibiotics (cefepime); fifth-generation cephalosporin antibiotics (ceftocycline); glycopeptide antibiotics (teicoplanin or vancomycin); macrolide antibiotics (azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin or spectinomycin); monobactam antibiotics (aztreonam); penicillin antibiotics (amoxicillin, Ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin, piperacillin, or ticarcillin); polypeptide antibiotics (bacitracin, colistin, or polymyxin b); quinolone antibiotics (ciprofloxacin, enoxacin, gatifloxacin, levofloxacin, lomefloxacin, moxifloxacin, norfloxacin, or ofloxacin); sulfonamide antibiotics (mafenamide, sulfazosulfonamide (old name), sulfacetamide, sulfadiazine, sulfamethoxazole, sulfadimethoxazole (old name), sulfasalazine, sulfadimethoxazole, trimethoprim, or trimethoprim -sulfamethoxazole (cotrimoxazole) (tmp-smx); tetracycline antibiotics (demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline); others (arsphenamine, chloramphenicol, clindamycin, lincomycin, ethambutol, fosfomycin, fusidic acid, furazolidone, isoniazid, linezolid, metronidazole, mupirocin, nitrofurantoin, pyrazinamide, quinupristin/dalfopristin, rifampicin (USA), thiamphenicol, metronidazole, dapsone, clofazimine); or cyclic lipopeptide antibiotics (daptomycin); glycylcycline antibiotics (tigecycline), or oxazolidinone antibiotics (linezolid).

In other embodiments, the at least one other therapeutic agent is an antifungal agent, such as a polyene antifungal agent (natamycin, chastomycin, filipinostatin, nystatin, amphotericin B, candesin); an imidazole antifungal agent (miconazole, ketoconazole, clotrimazole, econazole, bifonazole, butoconazole, fenticonazole, isoconazole, oxiconazole, sertaconazole, sulconazole, or tioconazole); a triazole antifungal agent (fluconazole, itraconazole, isavuconazole, rifluconazole, posaconazole, voriconazole, or terconazole); a thiazole antifungal drugs (abafungin); allylamine antifungal drugs (terbinafine, amorolfine, naftifine or butenafine); echinocandin antifungal drugs (anidulafungin, caspofungin or micafungin); others (benzoic acid, ciclopirox, tolnaftate, undecylenic acid, flucytosine or 5-flucytosine, griseofulvin, haloprogin and sodium bicarbonate); or substitutes (garlic, tea tree oil, citronella oil, iodine, lemongrass, olive leaf, orange oil, palmarosa oil, patchouli, lemon myrtle, neem seed oil, coconut oil, zinc or selenium). Or the drug is selected from any one of the drugs described in the present invention.

Compositions useful according to the invention can be evaluated for their efficacy in vitro and in vitro. See examples below. Briefly, in one embodiment, the ability of the composition, for example, to inhibit tumor cell proliferation in vitro can be tested. For in vivo studies, the composition can be fed to animals (e.g., mice) or injected or topically applied, and then evaluated for its effect on skin cancer cell survival, proliferation, metastasis, or one or more symptoms of skin cancer or related diseases or disorders. Based on the results, the appropriate dosage range, frequency, and route of administration can be determined.

The compositions useful in the present invention can be used alone or in combination with one or more other anti-epithelial cancer drugs or one or more other therapeutic drugs. The anti-epithelial cancer drug or other therapeutic drug can be or include a medical device, a medical product, a medicament or a drug. The primary efficacy of the anti-epithelial cancer drug or other therapeutic drug is to effectively alleviate one or more tumor diseases or conditions or one or more symptoms of one or more tumor diseases or conditions, or otherwise benefit the individual. Preferred therapeutic drugs include therapeutic food factors, immunogenic or immunostimulants, wound healing agents, etc.

It will be appreciated that the other anti-epithelial cancer or therapeutic agents (food-based drugs and pharmaceutical agents) described above may also be used in the methods of the present invention, either separately, simultaneously or sequentially with the compositions useful in the present invention.

It should be understood that composition dosage, administration time and general dosage regimen are different because of individual, and this depends on various variables, such as the severity of individual symptoms, the type of disease treated, the mode of administration selected and individual age, sex and/or overall health status. However, as can be seen by conventional example, for compositions that can be used for the present invention, dosage is by about 1mg to about 5000mg/kg, 1mg to about 4000mg/kg, 1mg to about 3000mg/kg, 1mg to about 2000mg/kg, 1mg to about 1000mg/kg per kilogram of body weight every day, preferably about 5mg to about 100mg/kg per kilogram of body weight, preferably administered every day. In one embodiment, dosage is about 0.05mg to about 250mg per kilogram of body weight that can be used for compositions of the present invention.

In various embodiments, sufficient composition is administered per day to achieve delivery of about 0.001 mg to about 50 mg, about 0.001 mg to about 40 mg, about 0.001 mg to about 30 mg, about 0.001 mg to about 20 mg, about 0.001 mg to about 10 mg, about 0.001 mg to about 5 mg, about 0.001 mg to about 1 mg, about 0.001 mg to about 0.5 mg, about 0.001 mg to about 0.1 mg, or about 0.001 mg to about 0.05 mg of a compound of Formula (I) or a pharmaceutically acceptable salt or solvate thereof per kilogram of body weight.

It should be understood that administration may include a single dose, such as a daily single dose, or multiple discrete doses may be given as needed. It should be understood that, taking into account the technology and the present disclosure, one of ordinary skill in the art will be able to determine the effective dosage regimen (including dosage and administration time) for a given condition without undue experimentation.

The present invention also relates to a dietary, nutritional, cosmeceutical or oral pharmaceutical composition, comprising one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and optionally, propolis or a material containing propolis in combination with cyclodextrin; or consisting mainly of, or consisting of. In certain embodiments, the composition is mainly composed of about 1-99wt% of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and about 1-80wt% of cyclodextrin. For example, the composition is mainly composed of about 20-80wt% of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and about 20-80wt% of cyclodextrin. In another example, the composition is mainly composed of about 20-40wt% of propolis, propolis resin or propolis resin extracts enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, and about 60-80wt% of cyclodextrin.

Provided is a dietetic, nutritional, cosmeceutical or oral pharmaceutical composition, comprising propolis, propolis resin or propolis resin extracts rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or a material containing propolis, propolis resin or propolis resin extracts enclosed by cyclodextrin, or consisting mainly of, or consisting of. In certain embodiments, the composition is mainly composed of about 1-30wt% of propolis, propolis resin or propolis resin extracts rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof and about 70-99wt% of cyclodextrin. For example, the composition is mainly composed of about 10-25wt% of propolis, propolis resin or propolis resin extracts rich in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof and about 75-90wt% of cyclodextrin. In another example, the composition consists essentially of about 20-30 wt % propolis, propolis resin or propolis resin extract enriched in one or more compounds of Formula (I) or one or more pharmaceutically acceptable salts or solvates thereof and about 70-80 wt % cyclodextrin.

In one embodiment, the composition comprises propolis, propolis resin or propolis resin extract enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof, or a propolis fraction enriched in one or more compounds of formula (I) or one or more pharmaceutically acceptable salts or solvates thereof. In one embodiment, the composition comprises at least about 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90 or 99 wt% of propolis, propolis resin or propolis resin extract or propolis component, and an effective range can be selected from any of these values (e.g., about 1-25 wt%, 1-30 wt%, 5-30 wt%, 15-30 wt%, 20-30 wt%, 25-30 wt%, 10-50 wt%, 15-50 wt%, 40-99 wt%, 45-99 wt%, 50-99 wt%, 55-99 wt%, 60-99 wt%, 65-99 wt%, 70-99 wt%, 75-99 wt%, 80-99 wt%, 85-99 wt%, 90-99 wt% or 95-99 wt%).

In one embodiment, the composition includes cyclodextrin, such as α-cyclodextrin and/or γ-cyclodextrin. In one embodiment, the composition includes at least about 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85 or 99 wt% of cyclodextrin, and an effective range can be selected from any of these values (e.g., about 1-99 wt%, about 5-99 wt%, about 10-99 wt%, about 15-99 wt%, about 20-99 wt%, about 25-99wt%, about 30-99wt%, about 35-99wt%, about 40-99wt%, about 45-99wt%, about 50-99wt%, about 55-99wt%, about 60-99wt%, about 65-99wt%, about 70-99wt%, about 75-99wt%, about 80-99wt%, about 85-99wt%, about 90-99wt% or about 95-99wt%).

When used in combination with another anti-epithelial cancer drug or therapeutic agent, the composition useful in the present invention and the other anti-epithelial cancer drug or therapeutic agent can be administered simultaneously or continuously. Simultaneous administration includes administration of a single dosage form (all ingredients) or administration of separate dosage forms substantially simultaneously. Continuous administration includes administration according to different schedules, preferably, so that there is overlap in the time of providing the composition useful in the present invention and other therapeutic agents.

In addition, it is contemplated that the compositions of the invention may be formulated with other active ingredients that may be beneficial to the individual under particular circumstances. For example, therapeutic agents directed to aspects of the same or different disease processes may be used.

Exemplary anti-epithelial cancer compositions and methods of preparing such compositions will now be described with reference to the following examples.

Example

Example 1: Initial bioassay-guided fractionation of propolis tincture

This example describes the evaluation of the anti-gastrointestinal cancer activity of propolis fractions generated by preparative chromatography. This study performed a proliferation assay on the human colon adenocarcinoma cell line DLD-1.

Preparation of Propolis Tincture Components by Column Chromatography

Use the glass chromatographic column that is filled with Merck Lichroprep CI8 reverse phase stationary phase (16x4 cm) to separate, wherein stationary phase is washed with methanol (MeOH) (200ml) and balanced with 20% ethanol aqueous solution (EtOH) (500ml). The propolis tincture dry solid (5.446g) dissolved in EtOH (5ml) is loaded into the top of the chromatographic column with a piston pump. Elution with a stepwise gradient solution (250ml) consisting of 20%, 30%, 40%, 50%, 60%, 70%, 80% and 90% ethanol aqueous solution, followed by elution with two 100% EtOH gradient solutions, and then elution with 2-propanol (IPA), ethyl acetate (EtOAc), acetone and chloroform (CHCl3). The solvents in the different components are removed by vacuum on a rotary evaporator, and then freeze-dried overnight. The two 100% EtOH fractions and the remaining four less polar fractions (IPA, EtOAc, acetone and CHCl3) were combined for bioassay work due to their relatively low quality. The fractions are shown in Table 1 according to the percentage of ethanol used in the elution step from the column (e.g., 20%, 30%, 40%, 50%, 60%, 70%, 80% and 90% aqueous EtOH and 100% EtOH).

Table 1 Propolis fractionation quality and sample number

Materials and methods for anti-gastrointestinal cancer and anti-proliferative assays in DLD-1 human colon adenocarcinoma cells, KYSE-30 human esophageal squamous cell carcinoma cells, and NCI-N87 human gastric cancer cells

As shown in Table 1 above, the samples obtained by separation of the dried solid of propolis tincture were evaluated for their ability to regulate the activity and proliferation of human colorectal adenocarcinoma cells (DLD-1) according to the MTT analysis. The dried solid raw material of propolis tincture is also required for the bioassay. In addition to the unsupplemented cell control (negative control), the study also included the positive control 5-fluorouracil (5-FU). In the subsequent examples, the ability of the samples to regulate the survival and proliferation of human esophageal cancer cells (KYSE-30) and human gastric cancer cells (NCI-N87) was also evaluated by MTT analysis.

Description of test materials and test methods

After thawing, human gastrointestinal cancer cell lines DLD-1, KYSE-30, and NCI-N87 were added to the test sample and reference sample for culture. The cell culture conditions should meet the requirements of the cell supplier (ATCC). The culture was then subjected to MTT analysis to determine the effect of the sample on cell proliferation.

The method is based on the steps reported below:

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Shimada, Y, Imamura, M, Wagata, T, Yamaguchi, N, Tobe, T. (1992) Characterization of 21 newly established esophageal cancer cell lines. Cancer, 69: 277–284.

Sutter, AP, Hopfner, M, Huether, A, Maaser, K, Scherubi, H. (2006) Targeting epithelial growth factor receptor with erlotinib (Tarceva ^™ ) for the treatment of esophageal cancer. Int J Cancer 118:1814–1822

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Yang, Y, Zhou, Z, He, S, Fan, T, et al. (2012) Treatment of prostate cancer with (Galectin-3)-targeted HPMA copolymer-(G3-C12)-5-fluorouracil conjugate. Biomaterials. 33: 2260-2271.

Nakagawa, Y., Iinuma, M., Naoe, T., Nozawa, Y., and Akao, Y. (2007). Characteristic mechanisms of α-mangostin-induced cell death: caspase-independent apoptosis, mitochondrial release of endonuclease-G, and increased expression of miR-143 in human colorectal cancer DLD-1 cells. Bioorganic and Medicinal Chemistry 15: 5620-5628.

Minegaki, T., Takara, K., Hamaguchi, R., Tsujimoto, M., and Nishiguchi, K. (2013). Factors affecting the sensitivity of human esophageal cancer cell lines to 5-fluorouracil and cisplatin. Oncology Express 5: 427-434.

Nakamura, A., Nakajima, G., Okuyama, R., Kuramochi, H., Kondoh, Y., Kanemura, T., Takechi, T., Yamamoto, M., and Hayashi, K. (2014). Calcium folinate enhances 5-fluorouracil-induced cytotoxicity in 5-fluorouracil-resistant gastric cancer cells after upregulation of thymidylate synthase expression. Gastric Cancer 17: 188-195.

Tankiewicz-Kwedlo, A., Pawlak, D., Domaniewski, T., and Buczko, W. (2010). Effects of erythropoietin, 5-fluorouracil, and SN-38 on the growth of DLD-1 cells. Pharmacological Reports 62: 926-937.

Sample preparation

Working solutions were prepared by dissolving the test components in 15% ethanol (ETOH)/HBSS to a concentration of 2 mg/ml solid.

Experimental procedures

Characteristics of the test system

1. Human colon adenocarcinoma cells were obtained from ATCC (CCl-221, DLD-1).

2. Human gastric cancer cells were obtained from ATCC (CRL-5822, NCI-N87).

3. Human esophageal squamous cell carcinoma cells were obtained from Sigma Aldrich (ECACC, KYSE-30).

4. The DLD-1 cell culture medium obtained from GIBCO was Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 mg/ml streptomycin. The culture medium was stored at 4°C.

5. The culture medium for NCI-N87 cells obtained from Sigma was RPMI-1640 medium modified to contain 2 mm L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 4500 mg/L glucose, and 1500 mg/L sodium bicarbonate. The culture medium was stored at 4°C.

6. The KYSE-30 cell culture medium obtained from Sigma was Ham's F12:RPMI-1640 (50:50) medium modified to contain 2 mm L-glutamine. For complete medium 10% FBS, 100 U/ml penicillin and 2.5 μg/ml gentamicin were added. The culture medium was stored at 4°C.

7. Penicillin-Streptomycin solution obtained from Sigma (#P-0781) includes 10,000 units/mL penicillin, 10 mg/ml streptomycin in 0.9% NaCl and stored at -20°C.

8. Trypsin-EDTA solution obtained from Invitrogen (15400054) included 0.25% trypsin/EDTA.

9. Phosphate buffered saline (PBS) was prepared in-house.

10. Obtain Hanks Balanced Salt Solution (HBSS) from GIBCO (14185-052) and store at 4°C.

11. Fetal bovine serum was obtained from GIBCO (10091-148) and stored at -20°C.

12. Obtain 100 mg/vial of MTT reagent from SIGMA (M-2128), dissolve in PBS at 10 mg/ml and store at -20° C. Prepare 5 mg/ml MTT solution in PBS and store at 4° C. as working solution.

13. Preparation of MTT lysis buffer consisting of 10% sodium dodecyl sulfate (SDS)/45% dimethylformamide: Dissolve 20 g SDS in 100 mL double distilled water (DDW), add 90 mL dimethylformamide, adjust the pH to 4.7 with glacial acetic acid, and then dilute with DDW to a final volume of 200 mL.

14. 5-Fluorouracil (5-FU) was obtained from Sigma (F-6627). Working solutions in 15% ETOH/HBSS were prepared as needed and then diluted to the desired concentration prior to bioassay.

Medium preparation

The proliferation medium for each cell line was as described above. Each medium was prepared according to the manufacturer's instructions and supplemented with penicillin-streptomycin solution (10 ml per liter). 10% FBS was added before use.

Cell culture

1. Thaw each cell line.

2. After initial proliferation using the above medium (see medium preparation), the culture was subcultured using trypsin-EDTA as described below. The medium was removed, 5 ml of trypsin-EDTA solution was added, and cultured at 37°C for 5 minutes or until all cells were detached. Trypsin was neutralized by adding an equal amount of DMEM medium and the culture was centrifuged at 300g (1200rpm) at 4°C for 5 minutes.

3. Decant the supernatant and resuspend the cell pellet in a culture medium containing DMEM, FBS (10%), penicillin (100 units/ml), and streptomycin (100 μg/ml). Cultivate the cells at 37°C in 5% CO ₂ /95% air.

4. Once confluence is reached, detach the cells using trypsin-EDTA and centrifuge as described in step 2.

5. Discard the supernatant and resuspend the cells at ^1.0x104 cells/ml in DMEM and supplements as described in step 3.

6. Add 180 μl of cells (1800 cells/well) or culture medium to each well of three 96-well plates. Incubate the plates at 37°C in 5% CO ₂ /95% air for 48 hours, which is sufficient for cells to adhere.

7. Add 20 μl of sample or positive control to each well. For "medium" or "cells only" controls, add 20 μl of 15% ETOH/HBSS to each well. Each sample is evaluated in 3-6 replicates, and the controls on each of the three microtiter plates are evaluated in 3-9 replicates (combined into triplicates). Unless otherwise noted, the final concentration of the sample in each well is 200 μg/ml.

8. The total volume of each well is 200 μl.

9. Incubate the plates at 37°C in 5% CO ₂ /95% air for 19 h.

Cell proliferation assay

1. After the incubation, add 20 μl of MTT working solution (5 mg/ml) to all wells and incubate at 37°C in 5% CO ₂ /95% air for 3-4 hours.

2. During the incubation period, it was noted that in some sample wells, including the cell sample wells and the sample medium blank control wells, unexpectedly high color development occurred. After the incubation period, remove the supernatant from each well, gently wash each well twice with HBSS to remove the purple color, and then add MTT lysis buffer as described in step 3 below.

3. Then add 100 μl of MTT lysis buffer and incubate the plate overnight at 37°C in 5% CO ₂ /95% air. Centrifuge the plate at 1200 rpm for 10 minutes to pellet any remaining insoluble material. Transfer a 200 μl aliquot from each well to a fresh 96-well plate. Read the plate at 570 nm using a VersaMax microplate reader.

4. The results are expressed as the percentage of proliferation of cells cultured in the sample compared to cells cultured in the control sample alone. The blank reading is subtracted from all wells as the background reading.

result

Table 2 summarizes the effects of the control and test samples on the proliferation of DLD-1 cells of propolis fractions S#1-S#10 and comparative propolis dry solid S#11, where NC = negative control (cells only), PC = positive control (5-fluorouracil, tested at 7.50 ng/ml). In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean of the measured optical density; p is the probability value of making the measurement statistically significant by the Student's t-test, here taken as <0.05 (NS = no significance); inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 2 Effects of propolis component samples on DLD-1 cell proliferation

The most active propolis fractions were fractions S#1, S#2, S#5, and S#8. Propolis fractions S#1, S#2, and S#5 contain known propolis phenolics and flavonoids, so they were not studied further here. Additional studies were performed to identify compounds present in propolis fraction S#8, which HPLC showed did not contain any previously identified phenolic compounds. These studies are described in Example 2.

Example 2: Initial bioassay-guided fractionation of phenol-free fractions

This example describes further bioassay-guided fractionation of propolis fraction S#8 prior to compound identification. Four additional subfractions were generated, numbered S#28 to S#31. As described in Example 1, this study performed a proliferation assay of the colon adenocarcinoma cell line DLD-1 by MTT assay.

Subfractions S#28 to S#31 were generated by preparative HPLC

The 90% ethanol aqueous solution elution component (propolis component S#8) generated in Example 1 was further fractionated by preparative HPLC. Propolis component S#8 was dissolved in pure ethanol and then chromatographically separated by preparative HPLC on a Phenomenex Synergi C-12C-12 column (4μ,-RP Max 80A 250x30 mm) using a Gilson 321 preparative pump and an Agilent 1100 series diode array detector. The injection volume was 0.5-1.5ml. The flow rate was 20ml/min. The solvent was 80% methanol aqueous solution (containing 0.1% TFA) and EtOAc/methanol (4:1vol/vol). The initial eluent component consisted of 80% methanol aqueous solution. Before the EtOAc/MeOH solvent concentration was linearly increased to 100% after 35 minutes, the solvent composition was kept at the initial conditions for 5 minutes. Chromatographic analysis was performed at room temperature (18-20°C). The components were collected manually and online detection and evaporative light scattering detection (ELSD) were performed at 210nm, 268nm and 327nm, respectively. The main component of this component is a weak polar component, which is then eluted in the chromatogram, and the minimum UV absorption appears at the wavelength commonly used to analyze other propolis components containing phenols, i.e. 268nm and 327nm. However, ELSD shows that the mixed components are more complex. Due to the destructiveness of ELSD, the preparation HPLC components were collected manually and online detection was performed at 210nm. Because the chromatographic analysis did not produce obvious isolated peaks, but rather a large rise and fall in the baseline, four components were collected during the experiment, and all components were prepared for bioassay.

Materials and methods for DLD-1 colon cancer antiproliferative assay

The ability of the samples shown in Table 3 to modulate the activity and proliferation of human colorectal adenocarcinoma cells (DLD-1) was evaluated according to the MTT assay. In addition to the unsupplemented cell control (NC, negative control), the study also included a positive control 5-fluorouracil (5-FU). Compared to Example 1, the concentration of 5-FU was increased to induce a stronger antiproliferative response. The DLD-1 antiproliferative assay was performed as described in Example 1.

Table 3 Samples

Sample No. Sample ID and concentration Sample No. Sample and concentration S#28 90％F1,200μg/ml NC-1 Cells only S#29 90％F2,200μg/ml PC-1 Cells + 5-FU 0.65μg/ml S#30 90％F3,200μg/ml PC-2 Cells + 5-FU 1.95 μg/ml S#31 90％F4,200μg/ml

The samples were dissolved in HBSS containing 15% ethanol as a 2 mg/ml working solution. When assayed, the final concentration of the samples was 200 μg/ml and the final EtOH concentration was 1.5%.

Results and discussion

Table 4 summarizes the effects of positive controls and samples on cell proliferation after 24 hours of culture, where NC = negative control (cells only), PC = positive control (5-fluorouracil, tested at 0.65 and 1.95 μg/ml, respectively). In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the measured mean optical density value; p is the probability value of making the measurement statistically significant by Student's t test, where the value is <0.05 (NS = no significance); inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 4 Effects of propolis subcomponent samples on DLD-1 cell proliferation

All propolis subfraction samples tested inhibited the proliferation of DLD-1 colon cancer cells. The most active fraction was S#30 90% F3, which completely inhibited proliferation but was cytotoxic at the concentrations tested. Further work was performed in Example 3 to identify the nature of the bioactive compounds of the samples used in Examples 1 (propolis fraction S#8) and 2 (propolis subfraction S#30).

Example 3: Identification of glycerides in propolis as active compounds

This example describes the separation and identification of novel dihydroxy fatty acid glycerides from NZ propolis resin fractions equivalent to Examples 1 (propolis fraction S#8) and 2 (propolis subfraction S#30). 1 g of 40% propolis tincture containing 40% propolis resin (400 mg dry solids) was fractionated on a silica column with a stepwise solvent gradient to produce 14 fractions with weight distributions as shown in Table 5. The fractions were analyzed by UHPLC using UV (detection of phenols at 268 nm) and ELSD detectors (for detection of all compounds). Propolis resin and selected fractions were also analyzed by LC-MS for structural elucidation. The purified subfractions were also analyzed by NMR. The column was washed sequentially with hexane, hexane/diethyl ether, and diethyl ether to elute most of the phenolic materials, including flavonoids, caffeic acid esters, and ferulic acid esters, from the silica (fractions 1-2; 3-7; 8-9). A brown component remained on the silica. When ethyl acetate (fractions 11-12) was used, brown began to elute. Most of the remaining dark color was then eluted with acetone and methanol (fractions 13, 14). The mass distribution showed that most of the propolis mass was present in the light yellow diethyl ether fraction, but a large part of the total mass was eluted in the fractions eluted with ethyl acetate (fractions 11, 12). The water-soluble material was recovered in fractions 13, 14 or retained on the column.

Table 5 Weight and solvent gradient of propolis silica gel fractionation

Components Eluent Volume(ml) Weight(mg) Percentage of total elution volume (%) 1 Hexane 10 1 0.3 2 Hexane 10 <1 <0.3 3 20% ether/hexane 10 1 0.3 4 40% ether/hexane 10 2 0.6 5 40% ether/hexane 10 8 2.3 6 60% ether/hexane 10 10 2.8 7 60% ether/hexane 10 62 17.5 8 Diethyl ether 10 70 19.8 9 Diethyl ether 10 51 14.4 10 Dichloromethane 20 53 15.0 11 Ethyl acetate 20 47 13.3 12 Ethyl acetate 20 38 10.7 13 Acetone/Methanol 11 3.1 14 Methanol 3 0.8

Results and discussion

These new compounds appeared as peaks in the 34-44 minute region of the ELSD chromatograms of fractions 12 and 13. The sharp and well-resolved late peaks observed in the ELSD were able to be correlated with a group of peaks seen in the LC-MS analysis of native propolis and enriched fractions 12 and 13. These peaks had no UV absorption associated with them. Mass spectrometry (MS) showed that ideal peaks appeared in the positive ion mode, where M+1 and M+Na peaks were observed. The molecular weights of the major components of this group of compounds were 460, 474, 488 and 502, corresponding to a homologous series, whose components differed due to the methylene groups. All mass spectra showed common peaks at 159 and 117 atomic mass units (amu). In the negative ion mode, the compound showed a quasi-molecular ion peak at M+45 due to formate adducts. High-resolution MS gave the molecular formula C25H48O7 for the 460 molecular weight compound, as well as other methylene groups for other compounds in the series. The subfractions of the native fractions 12 and 13 were then fully acetylated to help separate the related compounds and provide more clear spectral data. The mass spectrum of the fully acetylated mixture showed that these compounds were essentially triacetates, with an increase in molecular weight of 3x42 atomic mass units.

The above initial chromatographic separation was repeated several times to produce more enriched fractions 11-13. Related compounds were also easily separated from these enriched fractions by silica gel chromatography to produce sufficient material for NMR analysis. The spectrum showed the presence of four acetate groups, including the single acetate present in some natural compounds. Substituted glycerol was present and the acid attached to the glycerol was also hydroxylated at position 3. The remainder of the molecule consisted of a fatty acid chain which also had a second hydroxyl group. The fatty acids were dihydroxy C20, C21 C22, or C23 of the main compounds. Therefore, these compounds were glycerides.

GC-MS analysis of the TMS-treated glyceride-rich fraction generated by reverse phase column chromatography showed that the natural mixture of hydroxylated fatty acid lipids in propolis is very complex. This fraction contains the known common fatty acids present in propolis (palmitic, oleic and stearic acids) as well as monoglycerides. The complexity was reduced by removal of the glycerol and acetyl moieties by alkaline hydrolysis to obtain four major and minor dihydroxy fatty acids. This fits the above structural type, but the specific position of the acetyl group was not determined. When combined with the position/stereoisomer range of the fatty acids and hydroxyl groups, a large number of analogs were generated. The main compounds identified by chromatographic separation followed by structural elucidation using MS and NMR were monoacylglycerides of dihydroxy C20, C21 C22 or C23 fatty acids (glycerol esterified at C1). The fatty acids in the monoglycerides are mainly straight-chain C20-C22. The fatty acids are hydroxylated at C3, with the second hydroxyl group separated from the first by at least one methylene group; and not on the terminal methyl group of the fatty acid. The glycerol moiety can be acetylated at one or both hydroxyl groups at C2 and C3. Further work was performed in Example 4 to determine the location of the acetyl group and the second hydroxyl group on the fatty acid.

Example 4: Preparation of glyceride concentrate and separation and purification of glycerides

In this example, the position of the second hydroxyl group on the fatty acid was determined, as well as the position of the acetyl group. Monoglyceride compounds were also isolated. A bulk extract was prepared using 1.1 kg of dewaxed propolis resin using ethyl acetate as solvent. The propolis resin was coarsely crushed and soaked in ethyl acetate for 2 hours at room temperature with mechanical stirring. The solvent containing the dissolved extract was filtered but not concentrated. The extract was first adsorbed onto a portion of silica gel and the ethyl acetate was evaporated using a rotary evaporator; after that, about 20% of the extract solution was chromatographed on silica gel. Next, the previously adsorbed solid was applied to the top of a large silica gel column, and the column was eluted with 1:1 hexane/ether and 8×150 ml fractions were collected, of which four 150 ml fractions were eluted again with ether and finally with ethyl acetate. The four ethyl acetate fractions collected (fractions #13-#16) were much darker in color than the previous fractions. The final fraction (#17) was collected after elution with acetone. The progress of the purification process was followed using silica TLC (phosphomolybdic acid in ethanol developed by heating) and LC-MS. The glyceride-enriched fractions generated by silica gel chromatography were then combined and analyzed by reverse phase C18 column chromatography using a mixture of acid-free water and ethanol to obtain a final glyceride-enriched fraction (GLY-conc). This process was repeated with more ethyl acetate extracts as required.

All components were analyzed by LC-MS, indicating that ethyl acetate components #13-#16 and acetone component #17 contained major glyceride compounds. These components were also rich in brown materials, consistent with Example 3. Components #13 and #14 and components #15 and #16 were merged separately. The components after the #13/#14 merger were used for structural identification, as shown below. About 100mg of the #13/#14 mixture was mixed with 4ml THF:MeOH:H2O and 55mg LiOH. Stirred at ice temperature for 30 minutes and then at room temperature for 48 hours. Acidified with acetic acid and extracted with chloroform for the reaction mixture. The dry extract was then esterified with diazomethane and subjected to silica gel chromatography (ethyl acetate/hexane). The components containing a main spot were collected (Rf was about 0.3, where EtOAc:hexane was 1:1, and phosphomolybdic acid was colored in ethanol by heating). After being treated with BSTFA, the purified sample was analyzed by GC-MS. The 9 free fatty acid peaks (numbered 4-12) generated by the hydrolysis of glycerides in the mass spectrum are in the same sequence. The main peak 4 has a characteristic peak at 383 (corresponding to the loss of methyl and TMSOH), while the subsequent main peaks 5 (397), 7 (411), 10 (425), 11 (439) and 12 (453) are C19 to C24 dihydroxy fatty acids (methyl ester TMS ether). Another feature is that there are fragment ions at m/z 301 and 247 on all peaks. These fragments, especially the 301 fragment, are unique to 3,8-dihydroxy acid TMS ether, which is obtained by saponification, methylation and sialylation of 3-acetoxy, 8-hydroxy fatty acid glycerides esterified at the C2 position on glycerol [Asai, T., Hara, N., Kobayashi, S., Kohshima, S., Fujimoto, Y. (2009). Acylglycerols (=glycerides) in the exudate of glandular hairs of Paulownia tomentosa leaves. Helvetica Chimica Acta. 92: 1473-1494]. Other possible dihydroxy acids 3,6, 3,7 and 3,9 have different fragmentation patterns. The dihydroxy acids in the glyceride component are therefore 3,8-dihydroxy fatty acids from C19 to C24. The relative peak sizes indicate that C20 and C22 chain lengths are the main compounds. It was also observed that the secondary peaks 6, 8, 9, 13 have almost the same MS fragmentation pattern as the larger peaks. Peak 9 is very similar to peak 10, and peak 12 is very similar to peak 13. These secondary peaks are identical versions of the C22 and C24 acids, i.e., methyl branches at the ends of the fatty acids. This is consistent with the NMR evidence for fully acetylated compounds, which shows only a small amount of methyl branches.

For the combined fraction #15/16, preparative HPLC was first performed, followed by Sephadex LH20 size exclusion chromatography to obtain two monoacetate compounds GLY-1Ac-1 and GLY-1Ac-2 (C20 and C21 dihydroxy fatty acid monoacetates, respectively). For component #17, preparative HPLC was first performed, followed by Sephadex LH20 size exclusion chromatography to obtain three non-acetate compounds GLY-0Ac-1, GLY-0Ac-2 and GLY-0Ac-3 (C20, C21 and C22 dihydroxy fatty acid monoglycerides, respectively). The separation process was followed by HPLC combined with ELSD, and the isolated individual compounds were then evaluated by HPLC, LC-MS and NMR, with a purity of >95%.

Example 5: Preparation of glyceride concentrate, free fatty acids, methyl esters, diacetates, peracetates and cyclodextrin complexes

As described by people such as Reis, the glyceride concentrate sample obtained in Example 4 is hydrolyzed to generate free fatty acids (FFA-conc) (Reis, M.G., De Faria, A.D., Do Amaral, M.D.C.E., Marsaioli, A.J. (2003). Oncidinol-a novel diglyceride from Ornithophora radicans Barb.Rodr. (Orchidaceae) floral oil. Tetrahedron Communications 44:8519-8523). About 100 mg of glyceride concentrate is mixed with 4 ml THF:MeOH:H2O and 55 mgLiOH. Stir at ice temperature for 30 minutes, then at room temperature for 48 hours. The reaction mixture is acidified with acetic acid and extracted with chloroform. Use silica gel chromatography (ethyl acetate/hexane) to purify the free fatty acid mixture.

The sample of the free fatty acid mixture of purifying is methylated, to generate dihydroxy fatty acid methyl ester (ME-conc).Methylate by adding diazomethane solution (Aldrich, trimethylsilylation diazomethane, 2.0M hexane solution) in 10mg free fatty acid mixture.After reacting 1 hour under room temperature, use acetic acid cancellation reactant, at N2 lower evaporate to dryness solution.Use silica gel chromatography (ethyl acetate/hexane) purifying methyl ester mixture.Collect the component (Rf is about 0.3, wherein EtOAc: hexane is 1:1, makes phosphomolybdic acid develop the color in ethanol by heating) that comprises a main spot.

Dihydroxy fatty acid glyceride diacetate (GLY-2Ac-1 mixture) was prepared by partial acetylation of glyceride concentrate (15 mg was placed in THF containing a catalytic amount of dimethylaminopyridine and stirred with 1 equivalent of acetic anhydride at room temperature overnight). The reaction mixture was purified to give one spot on TLC.

The partially purified glyceride fraction (about 150 mg) was dissolved in acetic anhydride (20 mL) and a small amount of dimethylaminopyridine was added to fully acetylate the glyceride-rich fraction (PerAc-conc). After stirring overnight at room temperature, methanol and toluene (about 20 mL each) were added and evaporated to dryness on a rotary evaporator to make the reaction stronger. The residue was chromatographed on silica gel with a hexane/ethyl acetate mixture to obtain a purified peracetic ester mixture (about 80 mg).

The cyclodextrin complex of the glyceride concentrate obtained in Example 4 was prepared as follows. About 400 mg of glyceride concentrate was accurately weighed and placed in a flask. Then 1.2 g of ethanol was added to the vial and stirred to dissolve the solid to obtain a 25% ethanol solution of glyceride (by mass). The glycerides dissolved rapidly in ethanol. About 300 mg of α, β and γ cyclodextrin were accurately weighed respectively and placed in a mortar. Then about 400 mg of 25% glyceride tincture was added to each mortar containing one cyclodextrin using a calibrated syringe. The contents of each mortar were then mixed into a uniform paste using a pestle. Then excess water was added to each mortar and the contents were mixed until the complex was evenly dispersed in the water and was light yellow. The contents of each mortar were added to a weighed round-bottom flask. The contents of the flask were frozen by rotating the flask in a dry ice/acetone mixture. The contents of the flask were then freeze-dried. 370 mg of γ-cyclodextrin glyceride complex (g-CD glyceride concentrate), 390 mg of β-cyclodextrin glyceride complex (β-CD glyceride concentrate), and 360 mg of α-cyclodextrin glyceride complex (α-CD glyceride concentrate) were recovered from the flask.

Example 6: Antiproliferative activity of dihydroxy fatty acid glycerides on human skin cancer cell line: melanoma cell line A-2058

This example demonstrates that a number of glyceride compounds in NZ propolis have novel anti-skin cancer activity against the human melanoma cell line A-2508. This example also describes the general in vitro bioassay method used to determine the antiproliferative activity of the purified fractions and highly purified glycerides against human skin cancer cell lines and the activity of these test compounds against the human melanoma cell line A-2058. The three human skin cancer cell lines are:

1) Human melanoma cell line (A-2058).

2) Human epidermoid (squamous) carcinoma cell line (A-431).

3) Human basal cell carcinoma cell line (TE 354.T).

After thawing, the cell lines were cultured with the test and reference samples. The cultures were then subjected to MTT assay to determine the effect of the samples on cell viability and proliferation. The standard error of the mean was evaluated as a percentage; if SEM%>15, extreme outliers were removed. Preliminary significant differences (regardless of the presence or absence of outliers) were evaluated using an independent Student's t-test when α≤0.05. The method is based on the steps reported below:

Ahn, NG, Campbell JS (1993). Metabolic labeling of mitogen-activated protein kinases in A431 cells demonstrates phosphorylation on serine and threonine residues. Proc. Natl. Acad. Sci. USA 90: 5143–5147.

Galan-Cobo, A et al. (2014). Functional inhibition of aquaporin-3 by gold-based compounds leads to blockade of cell proliferation. Journal of Cellular Physiology. 229: 1787–1801.

Roomi, MW, Kalinovsky, J, et al. (2013) Effects of nutrient medium on matrix metalloproteinase-9 dimers in various human cancer cell lines. Int J Oncol. 44: 936–942.

Huang, HC, Lin, MK, et al. (2013). Cardenolide and bufadienolide from Euphorbia obesa and their cytotoxicity evaluation. Planta Medica. 79: 1362–1369.

Tilli, CMLJ, Stavast-Kooy, AJW, et al. (2003). Ajoene, an organosulfur component derived from garlic, reduces basal cell carcinoma tumor size by inducing apoptosis. Archives of Dermatology. 295:117–123.

Sample preparation

Unless otherwise stated, the samples were dissolved in 15% ethanol (EtOH)/HBSS to make working solutions and then diluted 10-fold to give a final concentration of 50 μg/ml in cells.

Experimental Procedure

Characteristics of the test system

1. Human melanoma cell line (A-2058, ATCC CRL 11147) was obtained from ATCC, Bethesda, Maryland, USA.

2. The human squamous (epidermoid) carcinoma cell line (A-431, ATCC CRL1555) was obtained from ATCC, Bethesda, Maryland, USA.

3. Human basal cell carcinoma cell line (TE 354.T, ATCC CRL7762) was obtained from ATCC, Bethesda, Maryland, USA.

4. Obtain Dulbecco's modified Eagle's medium (DMEM) from GIBCO (12100-038). Add 10% fetal bovine serum (FBS), 100 U/ml penicillin and 100 mg/ml streptomycin before use.

5. Penicillin-Streptomycin solution obtained from Sigma (#P-0781) includes 10,000 units/mL penicillin, 10 mg/ml streptomycin in 0.9% NaCl and stored at -20°C.

6. Trypsin-EDTA solution obtained from Invitrogen (15400054) included 0.25% trypsin/EDTA.

7. Phosphate buffered saline (PBS) was prepared in-house.

8. Obtain Hanks Balanced Salt Solution (HBSS) from GIBCO (#14185-052) and store at 4°C.

9. Fetal bovine serum was obtained from GIBCO (#10091-148) and stored at -20°C.

10. Obtain 100 mg/vial of MTT reagent from Sigma (M-2128), dissolve in PBS at 10 mg/ml and store at -20° C. Prepare 5 mg/ml MTT solution in PBS and store at 4° C. as working solution.

11. Preparation of MTT lysis buffer consisting of 10% sodium dodecyl sulfate (SDS)/45% dimethylformamide: Dissolve 20 g SDS in 100 mL double distilled water (DDW), then add 90 mL dimethylformamide to the solution. Adjust pH to 4.7 with glacial acetic acid, then dilute with DDW to a final volume of 200 mL.

12. 5-Fluorouracil (5-FU) was obtained from Sigma (F-6627). Three working solutions of 19.5 μg/ml, 6.5 μg/ml and 1.95 μg/ml were prepared and dissolved in 15% Ethanol/HBSS. The final concentrations were 1.95 μg/ml (15 μM), 0.65 μg/ml (5 μM) and 0.195 μg/ml (1.5 μM), respectively.

Medium preparation

The culture conditions of the cells were as described by the cell supplier (ATCC). As described above, the proliferation medium for each cell line was DMEM. The culture medium was prepared according to the manufacturer's instructions and supplemented with penicillin-streptomycin solution (10 ml per liter). 10% FBS was added before use.

Cell culture

1. Thaw various cell lines obtained from the American Type Culture Collection.

2. After the initial propagation using the above DMEM medium, the culture was subcultured with trypsin-EDTA. The medium was removed, 5 ml of trypsin-EDTA solution was added, and incubated at 37°C for 5 minutes or until all cells were detached. Trypsin was neutralized by adding an equal amount of DMEM medium and the suspension was centrifuged at 300g (1200rpm) at 4°C for 5 minutes.

3. Decant the supernatant and resuspend the cell pellet in DMEM containing FBS (10%), penicillin (100 units/ml), and streptomycin (100 μg/ml).

4. Once confluence is reached, detach the cells using trypsin-EDTA and centrifuge as described in step 2.

5. Discard the supernatant and resuspend the cells in DMEM and supplements at different cell concentrations per ml as described in step 3. The cell concentration of A-431 culture was increased from ^1x104 cells/ml to ^5x104 cells/ml, and the concentration of the other two cell lines (A2058 and TE 354) was increased to ^2x104 cells/ml.

6. For the three cell lines, add 180 μl of cells or culture medium to each well of a 96-well plate. Incubate the plates for different times at 37°C with 5% CO ₂ /95% air to allow cells to adhere. For A-431 cultures, the pre-incubation time was 3 hours, and for the other two cell lines (A2058 and TE 354.T), the pre-incubation time was 24 hours.

7. Add 20 μl of test compound or 5-FU to each well. Add 20 μl of 15% ETOH/HBSS to the wells labeled "Media" or "Cells Only". Record the number of evaluations for each test sample or control in the table in the example.

8. The total volume of each well is 200 μl.

9. Incubate the plates at 37°C in 5% CO ₂ /95% air for 24 h.

Cell proliferation assay

1. After the incubation, add 20 μl of MTT working solution (5 mg/ml) to all wells and incubate the plate at 37°C in 5% CO2/95% air for 3-4 hours. Monitor the plate every 30-60 minutes. If a few cells show the presence of crystals, add lysis buffer as in step 2.

2. Then add 100 μl of MTT lysis buffer and incubate the plate overnight at 37°C in 5% CO2/95% air. Centrifuge the wells at 300g (1200 rpm) for 10 minutes to pellet any remaining insoluble material. Transfer 200 μl aliquots from each well to a fresh 96-well plate. Read the plate at 550 nm using a VersaMax microplate reader.

3. The results are expressed as the percentage of adsorption of cells cultured in the sample compared to cells cultured in the control sample alone. The blank reading is subtracted from all wells as the background reading.

The glyceride samples GLY-1Ac-1 and GLY-1Ac-2 (dihydroxyglycerides acetylated at the C3 position of glycerol, C20 and C21 fatty acids, respectively) and GLY-0Ac-1, GLY-0Ac-2 and GLY-0Ac-3 (dihydroxyglycerides without acetate, C20, C21 and C22 fatty acids, respectively) were tested at a final concentration of 50 μg/ml and compared with the negative control NC-1 (cells only), the positive controls PC-1, PC-2, PC-3 (composed of 5-FU, tested at concentrations of 0.195, 0.65 and 1.95 μg/ml, respectively) and PC-4 (composed of monopalmitin (MGP)), which were tested at a concentration of 50 μg/ml.

Results and discussion

This example shows that many glyceride compounds in NZ propolis have novel anti-skin cancer activity against human melanoma cell line A-2508. The results of the bioassay are shown in Table 6. In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean of the measured optical density; p is the probability value for making the measurement statistically significant by Student's t-test, here taken as <0.05; inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 6: Antiproliferative activity of dihydroxy fatty acid glycerides against human melanoma cell line A-2508

Sample number and number of replicates, n Average value (OD 570nm) SEM P value (<0.05) Inhibition rate (%) NC-1 cells only, n=6 0.1260 0.0046 1 0.00 PC-1 cells + 5-FU, 0.195 μg/ml, n = 6 0.1339 0.0067 NS 0 PC-2 cells + 5-FU, 0.65 μg/ml, n = 6 0.1187 0.0043 NS 5.78 PC-3 cells + 5-FU, 1.95 μg/ml, n = 6 0.1383 0.0045 NS 0 PC-4 cells + MGP, 50 μg/ml, n = 3 0.0548 0.0016 1.7E-05 56.5 S#12 cells + GLY-1Ac-1, 50 μg/ml, n = 2 0.0249 0.0037 2.4E-05 80.2 S#13 cells + GLY-1Ac-2, 50 μg/ml, n = 2 0.0236 0.0006 1.4E-06 81.3 S#14 cells + GLY-0Ac-1, 50 μg/ml, n=2 0.0157 0.0001 1.3E-05 87.5 S#15 cells + GLY-0Ac-2, 50 μg/ml, n=2 0.0106 0.0001 9.7E-06 91.6 S#16 cells + GLY-0Ac-3, 50 μg/ml, n=2 0.0358 0.0055 5.2E-05 71.6

All highly purified compounds inhibited the proliferation of human melanoma skin cancer cell lines by 72-92% when tested at 50 μg/ml. The most potent of these compounds was the non-acetylated dihydroxyglycerol GLY-0Ac-2 with a fatty acid chain length of 22 carbons. Also tested at 50 μg/ml, these dihydroxyglycerols had better antiproliferative activity than the positive control monoglycerol palmitate (MGP). 5-Fluorouracil had low and insignificant activity when tested at three concentrations, 0.195, 0.65 and 1.95 μg/ml.

Example 7: Antiproliferative activity of dihydroxy fatty acid glycerides against human epidermoid carcinoma A-431

This example shows that many glyceride compounds in NZ propolis have novel anti-skin cancer activity against human epidermoid carcinoma cell line A-431. The glyceride samples GLY-1Ac-1 (C20 dihydroxy fatty acid glyceride acetylated at the C3 position of glycerol) and GLY-0Ac-1 (C20 dihydroxy fatty acid glyceride without acetate on glycerol) were tested for antiproliferative activity against human epidermoid carcinoma cell line A-431 at a final concentration of 50 μg/ml. Cell proliferation assays were performed as described in Example 6. Comparison samples were compared with negative control NC-1 (cells only) and positive controls PC-1, PC-2, PC-3 (composed of 5-FU, tested at concentrations of 0.195, 0.65 and 1.95 μg/ml, respectively) and PC-4 (composed of glycerol monopalmitate (MGP), tested at a concentration of 50 μg/ml).

The results of the bioassay are shown in Table 7. In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean value of the measured optical density; p is the probability value for making the measurement statistically significant by the Student's t-test, where the value is <0.05 (N=no significance); % inhibition is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 7: Antiproliferative activity of dihydroxy fatty acid glycerides against human epidermoid carcinoma cell line A-431

Sample number and number of replicates, n Average value (OD 570nm) SEM P value (<0.05) Inhibition rate (%) NC-1 cells only, n=6 0.2621 0.0158 NS 0.00 PC-1 cells + 5-FU, 0.195 μg/ml, n = 6 0.2364 0.0064 NS 9.8 PC-2 cells + 5-FU, 0.65 μg/ml, n = 6 0.2416 0.0111 NS 7.8 PC-3 cells + 5-FU, 1.95 μg/ml, n = 6 0.2561 0.0086 NS 2.3 PC-4 cells + MGP, 50 μg/ml, n = 3 0.0360 0.0034 2.5E-05 86.3 S#12 cells + GLY-1Ac-1, 50 μg/ml, n = 3 0.0507 0.0037 3.3E-04 80.6 S#14 cells + GLY-0Ac-1, 50 μg/ml, n=3 0.0484 0.0025 3.6E-05 81.5

When tested at 50 μg/ml, the two monoglyceride compounds tested, GLY-1Ac-1 and GLY-0Ac-1, had almost identical levels of proliferation inhibition, 80.6% and 81.5%, respectively. This example shows that the amount of acetate in glycerol is not significant in terms of activity. Also when tested at 50 μg/ml, the antiproliferative activity of these dihydroxy fatty acid glycerides was similar to that of the positive control monoglycerol palmitate (MGP). When tested at three concentrations of 0.195, 0.65 and 1.95 μg/ml, 5-fluorouracil had low activity and was meaningless.

Example 8: Antiproliferative activity of dihydroxy fatty acid glycerides against human basal cell carcinoma TE 354.T

The glyceride samples GLY-1Ac-1 (C20 dihydroxy fatty acid glyceride acetylated at the C3 position of glycerol) and GLY-0Ac-1 (C20 dihydroxy fatty acid glyceride without acetate on glycerol) were tested for their antiproliferative activity against the human basal cell carcinoma cell line TE 345 at a final concentration of 50 μg/ml. The cell proliferation assay was performed as described in Example 6. The samples were compared to the negative control NC-1 (cells only) and the positive controls PC-1, PC-2, PC-3 (consisting of 5-FU, tested at concentrations of 0.195, 0.65 and 1.95 μg/ml, respectively) and PC-4 (consisting of glycerol monopalmitate, tested at a concentration of 50 μg/ml).

This example shows that many glyceride compounds in NZ propolis have novel anti-skin cancer activity against human basal carcinoma cell line TE 354. The results of the bioassay are described in Table 8. In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean of the measured optical density; p is the probability value for making the measurement statistically significant by Student's t-test, here taken as <0.05 (NS=no significance); inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 8: Antiproliferative activity of dihydroxyglycerides against human basal cell carcinoma cell line TE 354

Sample number and number of replicates, n Average value (OD 570nm) SEM P value (<0.05) Inhibition rate (%) NC-1 cells only, n=6 0.2696 0.0096 NS 0.00 PC-1 cells + 5-FU, 0.195 μg/ml, n = 6 0.2548 0.0051 NS 5.49 PC-2 cells + 5-FU, 0.65 μg/ml, n = 6 0.2656 0.0061 NS 1.49 PC-3 cells + 5-FU, 1.95 μg/ml, n = 6 0.2553 0.0070 NS 5.31 PC-4 cells + MGP, 50 μg/ml, n = 3 0.2277 0.0065 0.0247 15.6 S#12 cells + GLY-1Ac-1, 50 μg/ml, n = 3 0.1959 0.0037 1.3E-03 27.3 S#14 cells + GLY-0Ac-1, 50 μg/ml, n=3 0.1672 0.0236 1.7E-03 38.0

The two monoglyceride compounds tested, GLY-1Ac-1 and GLY-0Ac-1, had similar levels of proliferation inhibition at 27.3% and 38.0%, respectively, when tested at 50 μg/ml. This example shows that the absence of acetate on glycerol can enhance the bioactivity of this skin cancer cell line. Also tested at 50 μg/ml, the antiproliferative activity of these dihydroxy fatty acid glycerides was positively superior to the positive control monoglycerol palmitate (MGP). The activity of 5-fluorouracil was low and insignificant when tested at three concentrations of 0.195, 0.65 and 1.95 μg/ml, respectively.

Example 9: Antiproliferative activity of dihydroxyglycerides and their derivatives against human colon adenocarcinoma cell line DLD-1

In this example, the monoglycerides GLY-1Ac-1 and GLY-1Ac-2 (dihydroxy fatty acid glycerides acetylated at the C3 position of glycerol, C20 and C21 fatty acid chain lengths) and GLY-0Ac-1 and GLY-0Ac-2 (non-acetylated dihydroxy fatty acid glycerides, C20 and C21 fatty acid chain lengths) were tested for antiproliferative activity against the colon adenocarcinoma cell line DLD-1 at a final concentration of 50 μg/ml. In a second assay, GLY-1Ac-1 was retested at 50 and 25 μg/ml, respectively, to provide an indicative dose response. Cell proliferation assays were performed as described in Examples 1 and 2. The samples were compared to the negative control NC-1 (cells only) and the positive controls PC-1, PC-2 (composed of 5-FU, tested at concentrations of 0.65 and 1.95 μg/ml, respectively) and PC-3 (composed of monopalmitin (MGP), tested at a concentration of 50 μg/ml). Also included in the table are data for cells from the first and second bioassay runs, NC-1 and NC-2.

This example shows that many glyceride compounds in NZ propolis have novel anti-gastrointestinal cancer activity against human colon adenocarcinoma cell line DLD-1. The results of the bioassay are shown in Table 9. In the table, OD is the optical density measured at 570nm; SEM is the standard error associated with the measured optical density mean; p is the probability value of making the measurement statistically significant by Student's t-test, here taken as <0.05 (NS=no significance); inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data shows that the test compound has anti-cancer proliferation potential.

Table 9: Antiproliferative activity of dihydroxy fatty acid glycerides against human adenocarcinoma cell line DLD-1

All monoglycerides with one or no acetate attached to glycerol had a proliferation inhibition rate between 68-91% when tested at 50 μg/ml. The most potent of these compounds in the first assay was the monoacetylated dihydroxyglyceride GLY-1Ac-2. The antiproliferative activity of these dihydroxyglycerides was similar to or better than the positive control monopalmitin (MGP), also tested at 50 μg/ml. GLY-1Ac-1 was retested at 50 and 25 μg/ml. The antiproliferative activity was 94% at 50 μg/ml and only 13% at 25 μg/ml; therefore, the LD50 for this compound is between 25-50 μg/ml. The activity of 5-fluorouracil was low and insignificant when tested at 0.65 and 1.95 μg/ml.

Example 10: Antiproliferative activity of dihydroxyglycerol fatty acid mixtures, their derivatives and cyclodextrin complexes against human adenocarcinoma cell line DLD-1

This example shows the antiproliferative activity of purified glyceride fraction GLY-conc, purified glyceride derivatives including free fatty acids (FFA-conc), methyl esters (ME-conc) and fully acetylated glycerides (PerAc-conc), and cyclodextrin complexes of glycerides g-CD GLY-conc (γ-cyclodextrin-included glyceride concentrate) and α-CD GLY-conc (α-cyclodextrin-included glyceride concentrate) on the human colon cancer cell line DLD-1. These fractions and samples were prepared as described in Examples 4 and 5, and the antiproliferative activity of the glyceride concentrate on the colon adenocarcinoma cell line DLD-1 was tested at a final effective concentration of 50 μg/ml. The cyclodextrin-included glyceride sample containing 25% glyceride concentrate (by mass) was tested at a concentration of 200 μg/ml (equivalent to 50 μg/ml glyceride). The samples were compared with the negative control NC-1 (cells only) and the positive controls PC-1, PC-2, PC-3 (consisting of 5-FU, tested at concentrations of 6.5, 19.5 and 58.5 μg/ml, respectively) and PC-4 (consisting of glycerol monopalmitate, tested at a concentration of 50 μg/ml). The results of the bioassay are shown in Table 10. In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean optical density measured; p is the probability value of making the measurement statistically significant by the Student's t-test, where the value is <0.05 (NS = not significant); the inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 10: Antiproliferative activity of dihydroxyglyceride concentrates, their derivatives and complexes against human colon adenocarcinoma cell line DLD-1

This example shows that the isolated glyceride mixture from NZ propolis, derivatives of glycerides including free fatty acids, alkyl esters and peracetate esters and cyclodextrin-included glycerides have novel anti-gastrointestinal cancer activity against the human colon adenocarcinoma cell line DLD-1. The most potent substance tested was the monoglyceride concentrate, which inhibited proliferation by 82%. The most potent derivative and complex was the methyl ester, with an inhibition of 47%, followed by the alpha and gamma cyclodextrin complexes, with inhibitions of 33% and 28%, respectively. The peracetate ester and free fatty acids showed the lowest antiproliferative activity, at 19% and 10%, respectively. The concentrations tested for these derivatives may have been too low. 5-Fluorouracil had low and insignificant activity when tested at three concentrations, 6.5, 19.5 and 58.5 μg/ml.

Example 11: Antiproliferative activity of dihydroxy fatty acid glycerides on human gastric cancer cell line NCI-N87

In this example, the monoglycerides GLY-1Ac-1 and GLY-1Ac-2 (dihydroxy fatty acid glycerides acetylated at the C3 position of glycerol, C20, C21, C22 fatty acid chain lengths), GLY-0Ac-1, GLY-0Ac-2, GLY-0Ac-3 (non-acetylated dihydroxy fatty acid glycerides, C20, C21, C22 fatty acid chain lengths) and GLY-2Ac-1 mixture (dihydroxy fatty acid glycerides acetylated at the C2 and C3 positions of glycerol, mixture of dihydroxy fatty acid chain lengths) were tested for antiproliferative activity against the human gastric cancer cell line NCI-N87 at a final concentration of 50 μg/ml. At the same time, GLY-1Ac-1 was also tested at concentrations of 25 and 10 μg/ml, respectively, to provide an indicative dose response. Cell proliferation assays were performed as described in Examples 1 and 2. The samples were compared with the negative control NC-1 (cells only) and the positive controls PC-1, PC-2, PC-3 (composed of 5-FU, tested at concentrations of 6.5, 19.5 and 58.5 μg/ml, respectively) and PC-4 (composed of glycerol monopalmitate (MCP), tested at a concentration of 50 μg/ml). The results of the bioassay are shown in Table 11. In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean optical density measured; p is the probability value of making the measurement statistically significant by the Student's t-test, here taken as <0.05; inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 11: Antiproliferative activity of dihydroxy fatty acid glycerides against human gastric cancer cell line NCI-N87

Sample number and number of replicates, n Average value (OD 570nm) SEM P value (<0.05) Inhibition rate (%) NC-1 cells only, n=6 0.0864 0.0128 1.00 0.00 PC-1 cells + 5-FU, 6.5 μg/ml, n = 6 0.0693 0.0062 NS 19.83 PC-1 cells + 5-FU, 19.5 μg/ml, n = 6 0.0792 0.0067 NS 8.32 PC-1 cells + 5-FU, 58.5 μg/ml, n = 6 0.0793 0.0023 NS 8.22 PC-4 cells + MGP, 50 μg/ml, n = 6 0.0000 0.0000 N/A 100 S#1 cells+GLY-0Ac-1, 50μg/ml, n＝6 0.0000 0.0000 N/A 100.00 S#2 cells + GLY-0Ac-2, 50 μg/ml, n = 6 0.0000 0.0000 N/A 100.00 S#3 cells + GLY-0Ac-3, 50 μg/ml, n = 6 0.0000 0.0000 5.0E-05 99.97 S#4a cells + GLY-1Ac-1, 50 μg/ml, n = 6 0.0000 0.0000 N/A 100.00 S#4b cells + GLY-1Ac-1, 25 μg/ml, n = 2 0.0118 0.0016 1.9E-02 86.38 S#4c cells+GLY-1Ac-1, 10μg/ml, n＝6 0.0530 0.0041 3.2E-02 38.61 S#7 cells + GLY-1Ac-2, 50 μg/ml, n = 6 0.0000 0.0000 N/A 100.00 S#8 cells+GLY-2Ac-1mix, 50μg/ml, n＝4 0.0414 0.0042 2.5E-02 52.08

This example shows that a large number of glyceride compounds in NZ propolis have new and very effective anti-gastrointestinal cancer activity against the human gastric cancer cell line NCI-N87. When tested at 50μg/ml, all isolated non-acetate and monoacetate compounds were cytotoxic (proliferation inhibition rate was 100%). The diacetate glyceride compound mixture - GLY-2Ac-1 mixture had a very strong anti-proliferative effect at an inhibition rate of 52%, but was not cytotoxic. When the test concentration of the monoacetate compound GLY-1Ac-1 was reduced from 50μg/ml to 25μg/ml, the anti-proliferative activity also dropped to 86% (still very active). Further reducing the test concentration from 25μg/ml to 10μg/ml, the anti-proliferative activity dropped to 39%. Therefore, the LD50 is between 25μg/ml and 10μg/ml. When tested at three concentrations of 6.5, 19.5 and 58.5 μg/ml, 5-fluorouracil showed low activity and no significance; when tested at 6.5 μg/ml, it showed the highest activity. When tested at a concentration of 50 μg/ml, monopalmitoylglycerol (MGP) also showed cytotoxicity.

Example 12: Antiproliferative activity of dihydroxyglycerol fatty acid mixtures, derivatives and cyclodextrin complexes against human gastric cancer cell line NCI-N87

This example shows the antiproliferative activity of the purified glyceride fraction GLY-conc, the derivatives of purified glyceride, including free fatty acids (FFA-conc), methyl esters (ME-conc) and fully acetylated glyceride (PerAc-conc), and the cyclodextrin complexes of glyceride g-CD GLY-conc (γ-cyclodextrin-included glyceride concentrate) and α-CD GLY-conc (α-cyclodextrin-included glyceride concentrate) on the human gastric cancer cell line NCI-N87. These components and samples were prepared as described in Examples 4 and 5, and the antiproliferative activity of the glyceride concentrate on the gastric cancer cell line NCI-N87 was tested at a final effective concentration of 50 μg/ml. The cyclodextrin-included glyceride sample containing 25% glyceride concentrate (by mass) was tested at a concentration of 200 μg/ml (equivalent to 50 μg/ml glyceride). The samples were compared with the negative control NC-1 (cells only) and the positive controls PC-1, PC-2, PC-3 (consisting of 5-FU, tested at concentrations of 6.5, 19.5 and 58.5 μg/ml, respectively) and PC-4 (consisting of glycerol monopalmitate (MCP), tested at a concentration of 50 μg/ml). The results of the bioassay are shown in Table 12. In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean optical density measured; p is the probability value of making the measurement statistically significant by the Student's t-test, where the value is <0.05 (NS = not significant); the inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 12: Antiproliferative activity of dihydroxyglycerol concentrates, derivatives and complexes against human gastric cancer cell line NCI-N87

This example shows that the isolated glyceride mixtures from NZ propolis, derivatives, including free fatty acids, alkyl esters and full acetate esters, and cyclodextrin-included glycerides have novel anti-gastrointestinal cancer activity against the human gastric cancer cell line NCI-N87. The most effective substance tested was the monoglyceride concentrate, with an inhibition rate of 82% proliferation. The most effective derivatives and complexes were the methyl ester (70% inhibition), β-cyclodextrin complex (78% inhibition), α-cyclodextrin complex (68% inhibition) and γ-cyclodextrin complex (65% inhibition). The full acetate and free fatty acids had relatively low antiproliferative activities of 60% and 55%, respectively. The activity of 5-fluorouracil was low and insignificant when tested at three concentrations of 6.5, 19.5 and 58.5 μg/ml; the highest activity was found when tested at 6.5 μg/ml. Monopalmitin (MGP) was cytotoxic at a test concentration of 50 μg/ml.

Example 13: Antiproliferative activity of dihydroxyglycerides against human esophageal cancer cell line KYSE-30

In this example, the monoglycerides GLY-1Ac-1 and GLY-1Ac-2 (acetylated dihydroxy fatty acid glycerides at the C3 position of glycerol, C20, C21 fatty acid chain lengths), GLY-0Ac-1, GLY-0Ac-2, GLY-0Ac-3 (non-acetylated dihydroxy fatty acid glycerides, C20, C21, C22 fatty acid chain lengths) and GLY-2Ac-1 mixture (acetylated dihydroxy fatty acid glycerides at the C2 and C3 positions of glycerol, mixture of fatty acid chain lengths) were tested for antiproliferative activity against the human esophageal cancer cell line KYSE-30 at a final concentration of 50 μg/ml. GLY-1Ac-1 was also tested at concentrations of 25 μg/ml and 10 μg/ml to provide an indicative dose response. Cell proliferation assays were performed as described in Examples 1 and 2. The samples were compared with the negative control NC-1 (cells only) and the positive controls PC-1, PC-2, PC-3 (consisting of 5-FU, tested at concentrations of 6.5, 19.5 and 58.5 μg/ml, respectively) and PC-4 (consisting of monopalmitin (MCP), tested at a concentration of 50 μg/ml). The results of the bioassay are shown in Table 13. In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean of the measured optical density (NS = not significant); p is the probability value of making the measurement statistically significant by the Student's t-test, here taken as <0.05; inhibition % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 13: Antiproliferative activity of dihydroxyglycerides against human esophageal cancer cell line KYSE-30

This example shows that many glyceride compounds in NZ propolis have novel and potent anti-gastrointestinal cancer activity against the human esophageal cancer cell line KYSE-30. All isolated non-acetate and monoacetate compounds had moderate to strong anti-proliferative effects (34-86%) when tested at 50 μg/ml, with the most potent being GLY-1Ac-1. The diacetate compound mixture, the GLY-2Ac-1 mixture, had a moderate anti-proliferative effect at 25% inhibition. When the test concentration of the monoacetate compound GLY-1Ac-1 was reduced from 50 μg/ml to 25 μg/ml, the anti-proliferative activity also dropped to 50% (still very potent activity). Further reducing the test concentration from 25 μg/ml to 10 μg/ml, the anti-proliferative activity reached a statistically significant level of 9%. Therefore, the LD50 is about 25 μg/ml. At a test concentration of 58.5 μg/ml, 5-fluorouracil had a low but significant activity of 17%, and at concentrations as low as 6.5 and 19.5 μg/ml, the activity was insignificant. At a test concentration of 50 μg/ml, glyceryl monopalmitate (MGP) had a strong antiproliferative activity of 75%.

Example 14: Antiproliferative activity of dihydroxyglycerol fatty acid mixture and its derivatives on human esophageal squamous cell carcinoma cell line KYSE-30

This example shows the antiproliferative activity of purified glyceride fraction GLY-conc, purified glyceride derivatives, including free fatty acids (FFA-conc), methyl esters (ME-conc) and fully acetylated glycerides (PerAc-conc) on human esophageal squamous cell carcinoma cell line KYSE-30. These components and samples were prepared as described in Examples 4 and 5, and the antiproliferative activity of glyceride concentrates on esophageal squamous cell carcinoma cell line KYSE-30 was tested at a final effective concentration of 50 μg/ml. Compare the samples with negative control NC-1 (cells only) and positive controls PC-1, PC-2, PC-3 (composed of 5-FU, tested at concentrations of 6.5, 19.5 and 58.5 μg/ml, respectively) and PC-4 (composed of glycerol monopalmitate (MCP), tested at a concentration of 50 μg/ml). The bioassay results are shown in Table 14. In the table, OD is the optical density measured at 570 nm; SEM is the standard error associated with the mean optical density measured; p is the probability value for making the measurement statistically significant by Student's t-test, where the value is <0.05 (NS = not significant); inhibition rate % is the percentage of proliferation reduction compared to the negative control, and a large amount of data indicates that the test compound has anti-cancer proliferation potential.

Table 14: Antiproliferative activity of dihydroxyglyceride concentrates and their derivatives against human esophageal squamous cell carcinoma cell line KYSE-30

Sample number and number of replicates, n Average value (OD 570nm) SEM P value (<0.05) Inhibition rate (%) NC-1 cells only, n=6 0.1519 0.0072 1.000 0.00 PC-1 cells + 5-FU, 6.5 μg/ml, n = 6 0.1336 0.0083 NS 12.02 PC-2 cells + 5-FU, 19.5 μg/ml, n = 6 0.1407 0.0053 NS 7.34 PC-3 cells + 5-FU, 58.5 μg/ml, n = 6 0.1263 0.0064 2.4E-02 16.85 PC-4 cells + MGP, 50 μg/ml, n = 6 0.0385 0.0047 1.2E-07 74.68 S#9 cells+GLY-conc, 50μg/ml, n＝6 0.1137 0.0033 7.2E-04 44.03 S#10 cells+FFA-conc, 50μg/ml, n=6 0.0850 0.0039 1.0E-05 21.47 S#11 cells+ME-conc, 50μg/ml, n=6 0.1193 0.0028 1.8E-03 27.59 S#12 cells+PerAc-conc, 50μg/ml, n=6 0.1100 0.0028 2.9E-04 32.03

This example shows that a mixture of glycerides isolated from NZ propolis and their derivatives, including free fatty acids, alkyl esters and peracetate esters, have novel anti-gastrointestinal cancer activity against the human esophageal squamous cell carcinoma cell line KYSE-30. The most potent substance tested was the monoglyceride concentrate, with a proliferation inhibition rate of 44%. The most potent derivative was the peracetate ester, with an inhibition rate of 32%. The methyl ester and free fatty acids had lower antiproliferative activity, at 28% and 21%, respectively. 5-Fluorouracil had a lower but significant activity of 16.8% at a test concentration of 58.5 μg/ml, and the activity was insignificant at concentrations as low as 6.5 and 19.5 μg/ml. Glycerol monopalmitate (MGP) had a strong antiproliferative activity of 75% at a test concentration of 50 μg/ml.

Example 15: Isolation of dihydroxy fatty acid glycerides from poplar

This example shows that certain poplar species or hybrids are the source of dihydroxy fatty acid glycerides found in New Zealand propolis and are therefore an alternative plant source of these glycerides in propolis or chemical synthesis.

Poplar bud, shoot and leaf samples were collected from the Plant and Food Research Poplar Collection Station at Aokautere (Manawatu, New Zealand) and the Wellington Regional Council Nursery at the Akura Conservation Centre (Masterton, New Zealand) from September to February. Most samples were taken from short-stemmed trees because they are more easily accessible.

The samples were weighed and extracted with absolute ethanol at room temperature for approximately 2 hours. The samples were extracted as is, without maceration or sectioning. The extracts were dried and weighed, and samples of each extract were prepared for LC-MS analysis by preparing an ethanol solution at a concentration of 5 mg/ml. Leaf and flower samples were collected from leaves and flower buds of two Paulownia tomentosa trees and a mature willow tree in the Hutt Valley for comparative analysis. When extracting the material, the plant material was immersed in ethyl acetate to extract only the waxy outer layer of the material.

All samples were analyzed using the LC-MS method developed for propolis glycerides as described in Example 4. This analytical method is capable of detecting very low levels of novel dihydroxy fatty acid glycerides and other glycerides, if present.

result

The qualitative contents of dihydroxy fatty acid glycerides in young branches, leaves or buds of various poplar varieties are shown in Table 15.

Table 15: Qualitative content of glycerides in New Zealand poplar species

np = absent glycerides, ++ moderate glycerides, +++ high glycerides, ND = not determined.

The analysis showed that the dihydroxy fatty acid glycerides found in propolis were also present in many of the more commonly grown New Zealand poplar species analysed. The presence of glycerides in leaves, twigs and buds of many poplars appears to be commonly associated with the resins in these parts.

The fatty acid composition of the glycerides is substantially the same as that in propolis, indicating that poplar is the source of propolis glycerides. It is expected that monoglycerides can be isolated from poplar exudates in a similar manner to that used in Example 4. Poplar species such as 'Tasman' and 'Fraser' are typical examples of widely planted European and American poplars (also known as Canadian poplars and Carolina poplars), while hybrid poplars such as Selwyn are also common in New Zealand. In other species such as the Liaoyang x black poplar clone and the Chinese white poplar species 'Yunan', dihydroxy fatty acid glycerides are generally not present. Glycerides were not found in hybrids that were not hybridized with American black poplars. There was no such glyceride in willow leaf and bud samples, but a small amount of such glycerides was present in Paulownia, indicating that these plant species are not the source of propolis glycerides and are not suitable for separating the glyceride compounds described in the present invention.

Example 16: Chirality determination of novel glycerides

This example shows that a naturally occurring novel dihydroxy fatty acid glyceride has a 3R,8R stereochemistry. Chiral analysis of the alcohol group of an organic molecule is often performed by using chiral derivatization reagents. These reagents come in both R and S forms and usually contain an aromatic group; when attached to the alcohol group, the aromatic group is able to affect the 1H NMR chemical shift of adjacent groups by shielding. Changes in the adjacent group shifts of the separately prepared R and S derivatives indicate the chirality of the original alcohol. For the novel glycerides, the stereochemistry of the alcohol group of the 3,8-dihydroxy fatty acid is of greatest concern. Since the length of the fatty acid chain has no effect on the chemical shifts of the C-2, -3, and -8 protons, a mixture of natural glycerides was used in this work.

Preparation and 1H-NMR Analysis of R,S Diester

The natural glyceride mixture was hydrolyzed using LiOH as described in Example 4. The free fatty acid methyl esters were then prepared using diazomethane from the hydrolyzed glyceride mixture described in Example 4. R- and S-α-methoxyphenylacetic acid (MPA), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (EDC) and 4-dimethylaminopyridine (DMAP) were purchased from Sigma. The chiral ester derivatization method used was described by Freire et al. in 2005. 10 mg of the dihydroxy free fatty acid methyl ester mixture was dissolved in about 0.8 mL of CH ₂ Cl ₂ (dry) in a 5 mL reaction bottle. The solution was stirred at room temperature using a magnetic stirrer and 23 mg of R-α-methoxyphenylacetic acid, 10 μL of EDC and a catalytic amount of DMAP were added. The reaction solution was washed with water, 1 M HCl, sodium bicarbonate and water to complete the reaction. The organic layer was dried in vacuo and purified using a small silica gel column to obtain the diester product (about 8 mg). The S-diester was obtained in a similar manner. 1H NMR samples were dissolved in CDCl ₃ and run at 500 Hz. The product was then checked by 1H NMR.

Results and discussion

The signals of H-2, H-3 and H-8 were separated sufficiently for detailed analysis. It was first observed that the two products were single species by NMR. This situation only occurs when the original alcohol is a chiral alcohol, i.e., the C-3 and C-8 hydroxyls are stereospecific and are R or S to each other. The shift changes of these derivatives Δδ (S-R) are shown in Table 16. Based on comparisons with similar compounds in the literature (Tables 17 and 18), the results indicate that the stereochemical configuration of all glycerides is 3R, 8R.

Table 16: 1H NMR chemical shift differences between R and S MPA esters of 3,8-dihydroxy fatty acid methyl esters

hydrogen Delta dihydroxy fatty acid methyl ester δDi R fatty acid methyl ester δDi S fatty acid methyl ester Δδ(S-R) _Ha -C(2) 2.5 2.6 2.43 -0.17 H _b -C(2) 2.41 2.5 2.35 -0.15 H-3 4.0 5.22 5.15 -0.07 H-8 3.6 4.8 4.9 +0.1

Confirmation of stereochemistry by comparison with literature

The stereochemistry was then confirmed by comparing the Δδ(S-R) shift changes with relevant examples from the literature. Two examples from the literature that were useful for direct comparison and used similar derivatization reagents were 3,7-diacetoxydocosahexaenoic acid, also known as byrsonic acid (Reis et al., 2007), and 1-acetyl-2-(3,7-diacetoxy-eicosanoyl)-glycerol, also known as oncidinol (Reis et al., 2003). Hydrolysis of the byrsonic acid removed the acetoxy group to produce the 3,7-dihydroxy fatty acid. The 3,7-diester was prepared using the chiral derivatizing agent Mosher's acid (methoxy{trifluoromethyl}phenylacetic acid) and then examined by 1H NMR. The Δδ(S-R) shifts are shown in Table 17, which led to the name (3R,7R)-3,7-dihydroxydocosanoic acid by Reis et al. (2007).

Table 17: 1H NMR chemical shift differences between R and S Mosher esters of 3,7-dihydroxy fatty acid methyl esters derived from byrsonic acids

hydrogen Δδ(S-R) _Ha -C(2) -0.11 H _b -C(2) -0.13 H-3 -0.08 H-7 +0.07

Similarly, Reis et al. (2003) also hydrolyzed oncidinol to remove glycerol and acetoxy groups to obtain 3,7 dihydroxy fatty acids; the acid was derivatized into a diester using Mosher acid and the 3,7 diester was detected by 1H NMR. As shown in Table 18, the acid was named 3R,7R dihydroxy acid due to the change in Δδ(S-R).

Table 18: 1H NMR chemical shift differences between R and S Mosher esters of 3,7-dihydroxy fatty acid methyl esters derived from oncidinol

hydrogen Δδ(S-R) _Ha -C(2) -0.11 H _b -C(2) -0.12 H-3 -0.06 H-7 +0.08

For byrsonic acid and oncinidol, the results were very similar to those of the new glycerides in Table 16. The Δδ(S-R) values were negative for H-2 and H-3 and positive for the distal hydroxyl position H8, demonstrating that the stereochemical configuration of all glycerides was 3R, 8R.

E,Riguera R.通过1H NMR光谱预测伯和仲1，2-二醇的绝对立体化学构型：原理和应用.《化学》2005 19；11(19):5509-22).Freire F,Seco JM,

E, Riguera R. Prediction of the absolute stereochemistry of primary and secondary 1,2-diols by 1H NMR spectroscopy: principles and applications. Chemistry 2005 19; 11(19): 5509-22).

Reis, M.G., De Faria, A.D., Dos Santos, I.A., Amaral, M.D.C.E., Marsaioli, A.J. Byrsonic acids: clues to floral mimicry in oil-producing flowers and oil-collecting bees. Journal of Chemical Ecology. 2007, 33(7), pp.1421-1429

Reis, M.G., De Faria, A.D., Do Amaral, M.D.C.E., Marsaioli, A.J. Oncidinol - a novel diacylglycerol from the floral oil of Ornithophora radicans Barb.Rodr. (Orchidaceae). 2003. Tetrahedron Communications. 44(46), pp.8519-8523

Industrial Applicability

The anti-epithelial cancer composition of the present invention comprises propolis or its extract or component and cyclodextrin rich in the compound of formula (I), and also comprises separated or purified compounds derived from propolis or its components, which can be used in the medical and healthcare fields, such as medical devices, medical supplies and medicines, and compositions. Methods for treating epithelial cancer, skin cancer and its symptoms using these compositions have been applied in the medical field.

Claims (23)

1. A pharmaceutical composition comprising a therapeutically effective amount of at least one compound selected from any one or more of the following:

a) 3, 8-dihydroxyeicosanoic acid,

b) 1- (3, 8-dihydroxyeicosanoyl) glycerol,

c) 1- (3, 8-dihydroxyeicosanoyl) 2-acetoxyglycerol,

d) 1- (3, 8-dihydroxyeicosanoyl) 3-acetoxyglycerol,

e) 1- (3, 8-dihydroxyeicosanoyl) 2, 3-diacetoxy glycerol,

f) 1- (3, 8-diacetoxy eicosanoyl) 2, 3-diacetoxy glycerol,

h) 3, 8-dihydroxydi-undecanoic acid,

i) 1- (3, 8-dihydroxydi-undecanoyl) glycerol,

j) 1- (3, 8-dihydroxydi-undecanoyl) 2-acetoxy glycerol,

k) 1- (3, 8-dihydroxydi-undecanoyl) 3-acetoxyglycerol,

l) 1- (3, 8-dihydroxyundecanoyl) 2, 3-diacetoxy glycerol,

m) 1- (3, 8-diacetoxy-di-undecanoyl) 2, 3-diacetoxy-glycerol,

n) methyl 3, 8-dihydroxyundecanoate,

o) 3, 8-dihydroxybehenic acid,

p) 1- (3, 8-dihydroxybehenoyl) glycerol,

q) 1- (3, 8-dihydroxybehenoyl) 2-acetoxyglycerol,

r) 1- (3, 8-dihydroxybehenoyl) 3-acetoxyglycerol,

s) 1- (3, 8-dihydroxybehenoyl) 2, 3-diacetoxy glycerol,

t) 1- (3, 8-diacetoxy-behenoyl) 2, 3-diacetoxy-glycerol, and

u) methyl 3, 8-dihydroxybehenate,

or a pharmaceutically acceptable salt of any one of a) to f) and h) to u).

2. The pharmaceutical composition of claim 1, wherein the composition is formulated for oral administration.

3. The pharmaceutical composition of claim 2, wherein the composition is in the form of a powder, liquid, tablet, caplet, hard or soft capsule or lozenge.

4. The pharmaceutical composition of claim 1, wherein the composition is formulated for topical administration.

5. The pharmaceutical composition of claim 4, wherein the composition is in the form of a lotion, cream, paste or salve.

6. The pharmaceutical composition of claim 4, wherein the composition is in the form of an ointment.

7. At least one compound selected from any one or more of the group consisting of

a) 3, 8-dihydroxyeicosanoic acid,

b) 1- (3, 8-dihydroxyeicosanoyl) glycerol,

c) 1- (3, 8-dihydroxyeicosanoyl) 2-acetoxyglycerol,

d) 1- (3, 8-dihydroxyeicosanoyl) 3-acetoxyglycerol,

e) 1- (3, 8-dihydroxyeicosanoyl) 2, 3-diacetoxy glycerol,

f) 1- (3, 8-diacetoxy eicosanoyl) 2, 3-diacetoxy glycerol,

g) Methyl 3, 8-dihydroxyeicosanoate,

h) 3, 8-dihydroxydi-undecanoic acid,

i) 1- (3, 8-dihydroxydi-undecanoyl) glycerol,

j) 1- (3, 8-dihydroxydi-undecanoyl) 2-acetoxy glycerol,

k) 1- (3, 8-dihydroxydi-undecanoyl) 3-acetoxyglycerol,

l) 1- (3, 8-dihydroxyundecanoyl) 2, 3-diacetoxy glycerol,

m) 1- (3, 8-diacetoxy-di-undecanoyl) 2, 3-diacetoxy-glycerol,

n) methyl 3, 8-dihydroxyundecanoate,

o) 3, 8-dihydroxybehenic acid,

p) 1- (3, 8-dihydroxybehenoyl) glycerol,

q) 1- (3, 8-dihydroxybehenoyl) 2-acetoxyglycerol,

r) 1- (3, 8-dihydroxybehenoyl) 3-acetoxyglycerol,

s) 1- (3, 8-dihydroxybehenoyl) 2, 3-diacetoxy glycerol,

t) 1- (3, 8-diacetoxy-behenoyl) 2, 3-diacetoxy-glycerol, and

u) methyl 3, 8-dihydroxybehenate,

or a pharmaceutically acceptable salt of any one of a) to u)

Use for preparing a composition having the following uses: inhibiting epithelial neoplasia, epithelial tumor growth, epithelial tumor metastasis or treating or preventing epithelial cancer in a subject; inducing apoptosis of one or more tumor epithelial cells in an individual; increasing responsiveness of the individual to treatment of the epithelial cancer; increasing sensitivity of an individual's epithelial tumor to treatment of epithelial cancer; re-sensitizing one or more epithelial cancer cells in the individual that are resistant to the treatment; at least partially reversing the resistance of tumor cells in an individual having an epithelial cancer to treatment of the epithelial cancer; reversing, in whole or in part, resistance of an epithelial cancer patient to treatment of epithelial cancer; or re-sensitize one or more tumors of the epithelial cancer patient that are resistant to, or are expected to be resistant to, the epithelial cancer treatment.

8. The use of claim 7, wherein the at least one compound is used with at least one other therapeutic agent to prepare the composition.

9. The use of claim 7, wherein the subject is a human subject.

10. The use of any one of claims 7-9, wherein the composition is formulated for oral administration.

11. The use of claim 10, wherein the composition is in the form of a powder, liquid, tablet, caplet, hard capsule or soft capsule or lozenge.

12. The use of any one of claims 7-9, wherein the composition is formulated for topical administration.

13. The use according to claim 12, wherein the composition is in the form of a lotion, cream, paste or salve.

14. The use according to claim 12, wherein the composition is in the form of an ointment.

15. Use of a pharmaceutical composition according to any one of claims 1-6 in the manufacture of a medicament for: inhibiting epithelial neoplasia, epithelial tumor growth, epithelial tumor metastasis or treating or preventing epithelial cancer in a human subject; inducing apoptosis of one or more tumor epithelial cells in a human subject; increasing responsiveness of a human individual to treatment of epithelial cancer; increasing the sensitivity of an epithelial tumor in a human individual to treatment of an epithelial cancer; re-sensitizing one or more epithelial cancer cells in a human individual that are resistant to the treatment; at least partially reversing the resistance of tumor cells to treatment of an epithelial cancer in a human subject having the epithelial cancer; reversing, in whole or in part, resistance of an epithelial cancer patient to treatment of epithelial cancer; or re-sensitize one or more tumors of the epithelial cancer patient that are resistant to, or are expected to be resistant to, the epithelial cancer treatment.

16. The use of claim 15, wherein the medicament is for treating or preventing epithelial cancer.

17. A process for the isolation or purification of at least one compound or a mixture of compounds as defined in any one of claims 1 to 6 from propolis or poplar or an extract or exudate thereof, comprising the steps of

a. Providing poplar extract, poplar propolis or extract or exudate thereof, and

b. separating or purifying compounds from poplar, propolis or extracts or exudates thereof,

wherein the poplar extract, poplar propolis or extract or exudate thereof is from populus nigra or populus tremulosa.

18. The method of claim 17, wherein the populus deltoidea comprises a variety or interspecific hybrid thereof.

19. The method of claim 18, wherein the populus euphratica is a species selected from the group consisting of: selwyn, tasman, luisa Avanzo and Fraser.

20. A process for the isolation or purification of at least one compound or a mixture of compounds as defined in any one of claims 1 to 6 from propolis or poplar or an extract or exudate thereof, comprising the steps of

a. Providing poplar extract, poplar propolis or extract or exudate thereof, and

b. Separating or purifying compounds from poplar, propolis or extracts or exudates thereof,

wherein the poplar extract, poplar propolis or extract or exudate thereof is from a hybrid of populus americana.

21. The method of claim 20, wherein the hybrid of populus jaborandi comprises populus jaborandi x populus jaborandi.

22. A process for the isolation or purification of at least one compound or a mixture of compounds as defined in any one of claims 1 to 6 from propolis or poplar or an extract or exudate thereof, comprising the steps of

a. Providing poplar extract, poplar propolis or extract or exudate thereof, and

b. separating or purifying compounds from poplar, propolis or extracts or exudates thereof,

wherein the poplar extract, poplar propolis or extract or exudate thereof is derived from a hybrid of Europe and America poplar.

23. A pharmaceutical composition comprising one or more compounds as defined in any one of claims 1 to 6, wherein the amount and/or concentration of the one or more compounds is specified by an indication associated with the pharmaceutical composition.