HK1249068A1

HK1249068A1 - Therapeutic uses of berberine formulations

Info

Publication number: HK1249068A1
Application number: HK18108954.7A
Authority: HK
Inventors: 卡尔‧奥斯卡‧布朗三世; 卡爾‧奧斯卡‧布朗三世; 曾博远; 林伊银; 曾博遠; 蔡陈恩; 陈志光; 林伊銀; 蔡陳恩; 陳志光
Original assignee: 安成生物科技股份有限公司
Priority date: 2015-06-24
Filing date: 2016-06-24
Publication date: 2018-10-26
Also published as: KR20180020229A; RU2018102700A; JP2018518507A; BR112017027897A2; RU2018102700A3; EP3313520A1; EP3313520A4; TW201713334A; IL256400A; AU2016282800A1; MX2018000262A; CA2990237A1; WO2016210230A1; RU2733743C2; CN107921284A

Abstract

Uses of pharmaceutical compositions comprising berberine for treatment of dermatologic toxicities and other skin disorders.

Description

Therapeutic use of berberine preparations

Background

Red surface-related skin disorders (red affected skin disorders) with the same similarity of symptoms and possible pathological causes include rosacea, acne vulgaris, seborrheic dermatitis, photodermatitis, and contact dermatitis. These red-face related conditions can range from heat and sensitivity to flushing or burning to a strong degree of sensitivity. Patients with red-face related skin disorders often exhibit extreme sensitivity to environmental and topical factors. Steroid-induced rosacea-like dermatitis (or steroid rosacea) is a papular or pustular lesion (version) with erythema and edematous bases with or without telangiectasia, which is caused by prolonged application of topical steroids to the face or a rebound condition following discontinuation of the topical steroid.

Skin toxicity is a known adverse skin event associated with targeted therapy or immunotherapy and has similar symptoms and possible pathological causes with red-face related skin disorders. Targeted therapies such as Epidermal Growth Factor Receptor (EGFR) inhibitors, Multiple Tyrosine Kinase (MTK) inhibitors, MEK inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, protein kinase b (akt) inhibitors, BRAF inhibitors, HER2 inhibitors, multiple kinase angiogenesis inhibitors, mTOR inhibitors, ALK/c-met inhibitors, multiple kinase Abl inhibitors, BTK inhibitors, HDAC inhibitors, proteasome inhibitors, and Retinoid X Receptor (RXR) agonists; immunotherapy such as cancer vaccines, cytokine agents (e.g., granulocyte-macrophage colony stimulating factor (GM-CSF), interferon and interleukin-2 (IL-2)), cell therapy (e.g., Tumor Infiltrating Lymphocytes (TIL), T Cell Receptor (TCR) engineered Peripheral Blood Lymphocytes (PBL) and Chimeric Antigen Receptor (CAR) engineered PBL), immune checkpoint protein inhibitors (e.g., PD-1, PD-L1, CTLA-4, TIM-3, LAG-3, BTLA, VISTA and TIGIT), and immune checkpoint protein stimulators (e.g., CD28, ICOS, 4-1BB, OX40, BITR, CD27, TWEAKR, HVEM, TIM-1 and CD-40); or a combination of any of the above therapies may induce toxic reactions, including papulopustular rashes (papulopustular rash), maculopapular rashes (maculopapular rash), erythema, telangiectasia flushing, paronychia and fissures, hair changes, xerosis, mucositis, pruritus and hand and foot skin reactions, which occur in more than 90% of patients and may also be superinfected with bacteria such as Staphylococcus aureus (Wollenberg, Kroth et al, Current side effects of EGFR inhibitors-antiparance and management, Dtsmed Canchenschr 2010; Lacouture, Maitland et al, A deployed epidermal receptor related inflammatory-specific gradient, tissue promoter, tissue, etc., (iii) toxins of immunotherapy for the practioner, Journal of Clinical Oncology, Vol.33, 2015; dy and Alex A.Adjei, understating, recogning, and framing toxicities of Targeted Anticancer therapeutics, CA Cancer J Clin, Vol.63, 2013; the Undergrade Mechanisms and Clinical Management, scientific, 2013; JLARKN et al, Combined Nivollumab and Iplilimumab or Monotherapy in Untreated Melanoma, N.Engl.J.Med., Vol.373, 2015). This histopathological finding of skin toxicity shows that inflammation is often involved and leads to acneiform skin rash. Papulopustular rash was reported more frequently in EGFR inhibitors such as cetuximab (cetuximab) (83% of patients) and afatinib (90% of patients), and MEK inhibitors such as semetinib (93% of patients) and trametinib (80% of patients) therapy. Maculopapular is more commonly described with PI3K inhibitors such as BKM-120 (37% of patients), MK2206 (52% of patients) therapy, immune checkpoint protein inhibitors such as the anti-CTLA-4 inhibitor plepima (ipilimumab) (33% of patients), the anti-PD-1 inhibitor nivolumab (26% of patients), or a combination of plepima and nivolimumab (more than 40% of patients).

Berberine (native yellow 18, 5, 6-dihydro-9, 10-dimethoxybenzo (g) -1, 3-benzodioxole (5,6-a) quinolizinium) is an isoquinoline alkaloid present in herbs such as coptis (rhizome of coptis), phellodendron (phenodinon), Scutellaria (Scutellaria bailii), mahonia aquifolium (mahonia aquifolium) and berberis (berberis). Berberine and its derivatives have been found to have antimicrobial and antimalarial activity. It can act on various pathogens, such as fungi, yeasts, parasites, bacteria and viruses.

Berberine also has anti-inflammatory effects, but the exact mechanism is not clear.

U.S. patent No. 6,440,465 relates to topical skin formulations of glucosamine in an emollient matrix containing berberine for the treatment of psoriasis. U.S. patent publication No. 2005/0158404 relates to a nutritional product, dietary supplement, or pharmaceutical composition containing vitamin a, vitamin E, selenium, vitamin B6, zinc, chromium, and an herbal source of berberine for oral administration to treat acne. U.S. Pat. No. 6,974,799 relates to topical compositions comprising a tripeptide (N-palmitoyl-Gly-His-Lys) and a tetrapeptide (N-palmitoyl-Gly-Gln-Pro-Arg) for treating visible signs of aging including wrinkles, striae distomark, dark circles. The preparation may contain other ingredients, including berberine. In these inventions, berberine is included as one of many ingredients, and the concentration thereof is not specified.

U.S. patent publication 2004/0146539 relates to a topical health care composition with slimming and skin-toning anti-aging benefits that can be used to treat skin aging, skin wrinkles, skin exfoliating (skinning), acne, rosacea, and other skin problems. The compositions of the invention include an antimicrobial agent selected from several agents, including berberine. In these nutraceutical compositions, berberine is included as one of the ingredients and its concentration is not specified. There has been 10% Mahonia aquifolium cream (Relieva) containing 0.1% berberine^TMApollo Pharmaceutical Canada Inc) for the treatment of psoriasis.

U.S. patent publication 2012/0165357 discloses the use of berberine for the treatment of various red face related skin disorders, but does not disclose any specific formulation of bupiriine that is effective in treating a specific condition.

Thus, there remains a need to develop new effective methods for treating various red-face related skin disorders as well as skin toxicity induced by targeted therapy and/or immunotherapy.

Disclosure of Invention

The present invention provides pharmaceutical compositions for the treatment and/or prevention of red-face related skin disorders and skin toxicity induced by targeted therapy and/or immunotherapy. The formulations provided are cream-based (i.e., cream) formulations or gel-based formulations.

In particular, the present invention provides a pharmaceutical composition comprising berberine, wherein the composition is a cream formulation comprising an aqueous phase and an oily phase.

In one embodiment, a cream formulation is provided in which the concentration of berberine is between 0.01% and 10%, preferably between 0.01% and 0.3% w/w, more preferably between 0.1% and 0.2% w/w, even more preferably between 0.1% and 0.15% w/w, most preferably about 0.12% w/w.

Whenever an amount or concentration is described herein in the form of w/w, the weight of each ingredient is based on the total weight of the formulation, unless specifically stated otherwise.

The pharmaceutical composition of the present invention may further comprise a penetration enhancer.

In one embodiment, the permeation enhancer is an anionic permeation enhancer.

In another embodiment, the penetration enhancer comprises60 and glycerol.

In one embodiment, berberine is the only pharmaceutically active component of the provided formulation.

In one embodiment, the pH of the pharmaceutical composition of the present invention is between about 4 to about 7, more preferably about 5.5.

In a preferred embodiment, the present invention provides a pharmaceutical composition comprising berberine as the only pharmaceutically active component, wherein the concentration of berberine is between 0.1% and 0.2% w/w, wherein the composition is a cream formulation comprising an aqueous phase and an oily phase, wherein the composition comprises a penetration enhancer, a preservative and a stabilizer, and wherein the pH of the composition is between about 4 and about 7.

In an even more preferred embodiment, the present invention provides a pharmaceutical composition comprising berberine as the only pharmaceutically active component, wherein the concentration of berberine is about 0.12% w/w, wherein the composition is a cream formulation comprising an aqueous phase and an oily phase, wherein the composition comprises a penetration enhancer, a preservative and a stabilizer, and wherein the pH of the composition is about 5.5.

In another embodiment, the present invention provides a pharmaceutical composition comprising berberine, wherein the composition is a gel-based formulation, wherein the composition comprises an anionic penetration enhancer.

In a preferred embodiment, the anionic penetration enhancer comprises Sodium Dodecyl Sulfate (SDS).

In one embodiment, in the gel-based pharmaceutical composition provided by the present invention, about 90% of the berberine has an average particle size of less than 10 μm.

In another embodiment, the gel-based pharmaceutical composition provided herein, about 50% of the berberine have an average particle size of less than 4 μm.

In one embodiment, the concentration of berberine in the provided gel-based formulation is between 0.01% and 0.3% w/w, more preferably between 0.1% and 0.2% w/w, even more preferably between 0.1% and 0.15% w/w, and most preferably about 0.12% w/w.

The present invention also provides a method of treating a red-face related skin condition comprising administering to a patient in need thereof a pharmaceutically effective amount of a pharmaceutical composition of the present invention.

In one embodiment, the red-related skin condition is selected from the group consisting of: rosacea, acne vulgaris, seborrhea, photodermatitis, contact dermatitis, steroid-induced rosacea-like dermatitis, and Epidermal Growth Factor Receptor (EGFR) inhibitor-induced skin disorders.

The present invention also provides a method for the treatment and/or prevention of skin toxicity induced by targeted therapy and/or immunotherapy comprising administering to a patient in need thereof a pharmaceutically effective amount of berberine and/or a biologically equivalent analogue thereof.

In preferred embodiments, the targeted therapy comprises therapy by an EGFR inhibitor, MTK inhibitor, MEK inhibitor, PI3K inhibitor, AKT inhibitor, BRAF inhibitor, HER2 inhibitor, multi-kinase angiogenesis inhibitor, mTOR inhibitor, ALK/c-met inhibitor, multi-kinase Abl inhibitor, BTK inhibitor, HDAC inhibitor, proteasome inhibitor, and RXR agonist.

In preferred embodiments, the immunotherapy includes therapy by cancer vaccines, cytokine agents (e.g., granulocyte-macrophage colony stimulating factor (GM-CSF), interferon, and interleukin-2 (IL-2)), cell therapies (e.g., Tumor Infiltrating Lymphocytes (TIL), T Cell Receptor (TCR) -engineered Peripheral Blood Lymphocytes (PBL), and Chimeric Antigen Receptor (CAR) -engineered PBL), immune checkpoint protein inhibitors (e.g., PD-1, PD-L1, CTLA-4, TIM-3, LAG-3, BTLA, VISTA, and TIGIT), and immune checkpoint protein stimulators (e.g., CD28, ICOS, 4-1BB, OX40, BITR, CD27, TWEAKR, HVEM, TIM-1, and CD-40).

In a preferred embodiment, berberine and/or a biologically equivalent analogue thereof is administered to the patient in the form of a topical pharmaceutical composition.

Drawings

This patent or application document contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the office upon request and payment of the necessary fee.

FIG. 1 is a graph of cumulative permeation of small chaetomine (ng/cm) for six tested berberine formulations²) -a time map.

FIG. 2 is a cumulative permeation of small chaetomine (ng/cm) of three berberine gel suspension formulations (G22, G23, and G24)²) -a time map.

Fig. 3 is a graph showing hematoxylin and eosin (H & E) staining of bilateral skin biopsies of nasolabial folds of patients who received afatinib and topically applied a gel formulation on one side of their face (G23).

Fig. 4 is a graph showing hematoxylin and eosin (H & E) staining of bilateral skin biopsies of the nasolabial fold of patients who received the EGFR inhibitor afatinib and topically administered the vehicle gel (berberine-free G23) on the other side of their face.

Figure 5 is a plot of papule count versus time for patients receiving afatinib and topically applying a gel formulation (G23) on one side of their face and a vehicle gel (berberine-free G23) on the other side of their face (indicating P <0.05 by Wilcoxon Signed Rank test (Wilcoxon Signed Rank test).

Fig. 6 is a graph of pustule count versus time for patients receiving afatinib and topically applying gel formulation (G23) on one side of their face and vehicle gel (berberine-free G23) on the other side of their face (indicated by wilcoxson symbol rank test, P < 0.05).

Figure 7 is a plot of total counts (papules and pustules) -time for patients receiving afatinib and topically applying the gel formulation (G23) on one side of their face and the vehicle gel (berberine-free G23) on the other side of their face (x denotes by wilcoxson signed rank test, P < 0.05).

Detailed Description

Depending on the particular condition being treated, it is important that berberine effectively penetrate the skin. Berberine is a hydrophilic compound (partition coefficient of 1.07 in octanol-water system) that makes it difficult for berberine to penetrate through the Stratum Corneum (SC) to reach the target site, e.g. dermis or epidermis, where red-related skin disorders or targeted therapy-induced skin toxicity may occur. In addition, berberine is rather soluble (solubility of 1.57mg/ml) and will therefore be released rapidly into the target cells, leading to temporary effects.

Accordingly, the present invention provides pharmaceutical compositions with enhanced berberine permeability for the treatment and/or prevention of red-face related skin disorders and skin toxicity induced by targeted therapy and/or immunotherapy. The formulations provided are cream-based (i.e., cream) formulations or gel-based formulations.

Since the cream formulation of the present invention can promote the penetration of berberine into the skin, a relatively small amount of berberine is sufficient to achieve the desired therapeutic effect. In one embodiment, the concentration of berberine in the provided cream formulation is between 0.01% and 10% w/w, preferably between 0.01% and 0.3% w/w, more preferably between 0.1% and 0.2% w/w, even more preferably between 0.1% and 0.15% w/w, most preferably about 0.12% w/w, based on the total weight of the formulation.

In one embodiment, the permeation enhancer is an anionic permeation enhancer. For example, the anionic penetration enhancer may comprise Sodium Dodecyl Sulfate (SDS).

In another embodiment, the penetration enhancer comprises60 and glycerol. The cream formulation of the present invention preferably comprises60 and glycerol as permeation enhancers. When the same penetration enhancers were used in non-cream formulations, they did not result in an increase in penetration rate, indicating that there is some uniqueness with regard to cream-based formulations.

In one embodiment, berberine is the only pharmaceutically active component of the provided formulation. Even though a component of the provided formulation may be an active ingredient in prior art formulations for purposes other than treating skin toxicity induced by targeted therapy or immunotherapy, it would still be considered a pharmaceutical excipient for the purposes of the provided formulation, provided that the component is not present in an amount sufficient to effectively treat skin toxicity induced by targeted therapy or immunotherapy.

In one embodiment, the pH of the pharmaceutical composition of the present invention is from about 4 to about 7, more preferably about 5.5.

In a preferred embodiment, the present invention provides a pharmaceutical composition comprising berberine as the only pharmaceutically active ingredient, wherein the concentration of berberine is between 0.1% and 0.2% w/w, wherein the composition is a cream formulation comprising an aqueous phase and an oily phase, wherein the composition comprises a penetration enhancer, a preservative and a stabilizer, and wherein the pH of the composition is between about 4 and about 7.

In an even more preferred embodiment, the present invention provides a pharmaceutical composition comprising berberine as the only pharmaceutically active ingredient, wherein the concentration of berberine is about 0.12% w/w, wherein the composition is a cream formulation comprising an aqueous phase and an oily phase, wherein the composition comprises a penetration enhancer, a preservative and a stabilizer, and wherein the pH of the composition is about 5.5.

It was very surprising and unexpected to find that the cream formulation of the present invention has an excellent permeability compared to non-cream berberine formulations.

In a preferred embodiment, the anionic penetration enhancer comprises Sodium Dodecyl Sulfate (SDS). The inclusion of SDS as an anionic penetration enhancer results in the provided gel-based formulation being hydrophobic (partition coefficient of 50.1 in octanol-water system) and having a significantly lower solubility, about 0.011mg/ml, enabling sustained release of berberine into the target cells, resulting in an extended release profile.

In the present invention it was found that berberine solubility ranges from 0.01 to 0.06mg/ml in the presence of SDS at a pH between 4 and 7, i.e. 25 to 150 times lower than aqueous berberine solubility (1.57mg/ml) and relatively low at pH 5.5.

It was surprisingly found that the penetration enhancers (SDS, glycerol, propylene glycol, PEG 400, ethanol and) The addition of SDS to the gel-based formulation resulted in the greatest permeability enhancement and the greatest local increase in berberine concentration in the epidermis and dermis.

It was also surprisingly found that in gel-based formulations there is a positive correlation between the amount of SDS and the permeability and a negative correlation between the size of berberine and the permeability.

Since the gel-based formulation of the present invention can facilitate the penetration of berberine into the skin, a relatively small amount of berberine is sufficient to achieve the desired therapeutic effect. In one embodiment, the gel-based formulation is provided with a berberine concentration of 0.01% to 0.3% w/w, more preferably 0.1% to 0.2% w/w, even more preferably 0.1% and 0.15% w/w, most preferably about 0.12% w/w, based on the total weight of the formulation.

The present invention also provides a method of treating and/or preventing skin toxicity induced by targeted therapy and/or immunotherapy comprising administering to a patient in need thereof a pharmaceutically effective amount of a pharmaceutical composition of the present invention.

In one embodiment, the targeted therapy is selected from the group consisting of: EGFR, Multiple Tyrosine Kinase (MTK), MEK, phosphoinositide 3-kinase (PI3K), protein kinase b (akt), BRAF inhibitors, HER2 inhibitors, multiple kinase angiogenesis inhibitors, mTOR inhibitors, ALK/c-met inhibitors, multiple kinase Abl inhibitors, BTK inhibitors, HDAC inhibitors, proteasome inhibitors, and RXR agonists; the immunotherapy is selected from the group consisting of: cancer vaccines, cytokine agents (e.g., granulocyte-macrophage colony stimulating factor (GM-CSF), interferon and interleukin-2 (IL-2)), cell therapies (e.g., Tumor Infiltrating Lymphocytes (TIL), T Cell Receptor (TCR) engineered Peripheral Blood Lymphocytes (PBL) and Chimeric Antigen Receptor (CAR) engineered PBL), immune checkpoint protein inhibitors (e.g., PD-1, PD-L1, CTLA-4, TIM-3, LAG-3, BTLA, VISTA and TIGIT), and immune checkpoint protein stimulators (e.g., CD28, ICOS, 4-1BB, OX40, BITR, CD27, TWEAKR, HVEM, HV1 and CD-40); and the skin toxicity induced by targeted therapy and/or immunotherapy is selected from the group consisting of: papulopustular rash, maculopapular rash, erythema, telangiectasias flushing, paronychia and fissures, hair changes, xerosis, mucositis, pruritus and hand-foot skin reactions.

The concentration of berberine in the epidermis, dermis and receptor (which refers to the container filled with PBS in contact with the skin) was measured by the following method. Franz diffusion cell devices are basically a piece of skin sandwiched between two clamps. The drug is applied to one side of the skin (top) and the drug concentration is measured at the receiving portion of the device (bottom).

As used herein, the term "permeability" refers to the amount of berberine present per gram of epidermal or dermal tissue, or per cm present in the receiver, since the formulation was applied to the skin for a period of time²Amount of berberine in the skin.

The amount of drug measured in the receiver represents the total amount of SC, epidermal and dermal regions that permeate through the skin. The pharmaceutical composition of the present invention has an improved permeability, and the preferred ranges of permeability are as follows:

epidermis: 0.4-4000 mug berberine per gram tissue

Dermis: 0.003-30 mug berberine per gram tissue

A receiver: each 1X 1cm²Skin 0.0001 to 1 μ g berberine.

Table 1 below lists various ingredients that may be used in the compositions of the present invention. However, this list is provided for illustrative purposes only, and is not meant to limit the scope of the present invention. Furthermore, the different ingredients/excipients may function in more than one way, e.g. may act as penetration enhancers, emulsifiers, wetting agents, etc.

TABLE 1

As used herein, the term "berberine" refers to 5, 6-dihydro-9, 10-dimethoxybenzo (g) -1, 3-benzodioxole (5,6-a) quinolizinium. The present invention also contemplates the use of berberine analogues including, but not limited to jatrorrhizine (jatrorrhizine), palmatine (palmatine), coptisine, 9-demethylberberine, 9-demethylpalmatine, 13-hydroxyberberine, berberine (berberberrubine), palmatine (palmatine), 9-O-ethyl berberine, 9-O-ethyl-13-ethyl berberine, 13-methyl dihydroberberine N-methyl salt, tetrahydroprotoberberine and its N-methyl salt, 9-lauryl berberine chloride, and pharmaceutically acceptable salts of all of these compounds.

As used herein, the term "pharmaceutically acceptable salt" includes salts of acidic or basic groups. Examples of pharmaceutically acceptable salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, and the like, as well as salts derived from relatively nontoxic organic acids such as acetic acid; propionic acid; isobutyric acid; maleic acid; malonic acid; benzoic acid; succinic acid; suberic acid; fumaric acid; mandelic acid; phthalic acid; benzenesulfonic acid; toluene sulfonic acids including p-toluene sulfonic acid, m-toluene sulfonic acid and o-toluene sulfonic acid; citric acid; tartaric acid; methanesulfonic acid; and so on. Also included are salts of amino acids such as arginine and the like, and salts of organic acids such as glucuronic or galacturonic acids and the like.

As used herein, the term "treating" includes inhibiting a disease or condition, causing a reduction in the severity and/or frequency of symptoms, eliminating symptoms and/or underlying causes, preventing the occurrence of symptoms and/or underlying causes thereof, alleviating and/or improving the condition of a patient. Thus, "treating" a patient with a composition described herein includes preventing a particular disorder in a susceptible individual, as well as managing clinically symptomatic individuals to inhibit or cause regression of the disorder or disease, and to maintain the current state and/or prevent progression of the disorder or disease. Treatment may include prophylaxis, treatment or cure.

As used herein, the term "pharmaceutically effective amount" of a compound and/or pharmaceutical composition of the present invention refers to an amount of the compound and/or composition sufficient to treat, inhibit, alleviate, or prevent various red-face related skin disorders (including, but not limited to, targeted therapy-induced skin toxicity) at a reasonable benefit/risk ratio suitable for any medical treatment. However, it will be understood that the total daily amount of the compounds and/or compositions of the present invention will be determined by the attending physician within the scope of sound medical judgment. The specific effective dose level for any particular patient will depend upon a variety of factors including the condition being treated and the severity of the condition; the activity of the particular compound used; the specific composition used; the age, weight, general health, sex, and diet of the patient; time of administration, route of administration, and rate of excretion of the particular compound used; the duration of the treatment; drugs used in combination or concomitantly with the specific compound used; and similar factors well known in the medical arts. For example, it is well known to those skilled in the art to start doses of the compositions at levels below those required to achieve the desired therapeutic effect and to gradually increase the dose until the desired effect is achieved.

The pharmaceutical compositions may also include a pharmaceutically acceptable carrier and may be in solid or liquid form, including but not limited to tablets, powders, capsules, pills, solutions, suspensions, elixirs, emulsions, gels, creams, patches or suppositories, including rectal and urethral suppositories.

As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. The pharmaceutically acceptable carrier is compatible with the other ingredients of the composition, the mode of administration, and does not harm the patient. The pharmaceutically acceptable carrier may be aqueous or non-aqueous. Pharmaceutically acceptable carriers include gums, starches, sugars, cellulosic materials and mixtures thereof. Some examples of materials that can be used as pharmaceutically acceptable carriers include, but are not limited to: (a) sugars such as lactose, glucose and sucrose; (b) starches, such as corn starch and potato starch; (c) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (d) tragacanth powder; (e) malt; (f) gelatin; (g) talc powder; (h) excipients, such as cocoa butter and suppository waxes; (i) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (g) glycols, such as propylene glycol; (k) polyols such as glycerol, sorbitol, mannitol and polyethylene glycol; (l) Esters, such as ethyl oleate and ethyl laurate; (m) agar; (n) buffering agents such as magnesium hydroxide, aluminum hydroxide, boric acid and sodium borate and phosphate buffers; (o) alginic acid; (p) pyrogen-free water; (q) isotonic saline; (r) ringer's solution; (s) ethanol; (t) phosphate buffer solution; and (u) other non-toxic compatible materials suitable for use in pharmaceutical compositions.

The compositions of the present invention may be administered using any means known in the art, including but not limited to oral, nasal, parenteral, topical, transdermal or rectal routes of administration. Preferably, the composition is suitable for oral or topical administration. For example, the active ingredients of the compositions may be formulated with suitable excipients for the preparation of tablets, capsules, pills, troches (troche), dragees (lozenes), solutions, powders or granules, suspensions, hard or soft capsules, patches and any other suitable form.

The present invention also provides a method of treating and/or preventing skin toxicity induced by targeted therapy and/or immunotherapy comprising administering to a patient in need thereof a pharmaceutically effective amount of berberine or a biologically equivalent analogue thereof.

The present invention also provides a method of treating and/or preventing skin toxicity induced by targeted therapy and/or immunotherapy comprising topically applying to the affected skin a pharmaceutically effective amount of a topical pharmaceutical composition comprising berberine or a biologically equivalent analogue thereof.

In one embodiment, the topical pharmaceutical composition is in the form of a lotion, cream, ointment, paste, gel, spray, suspension, emulsion, foam, patch, powder, and liniment.

In one embodiment, the topical pharmaceutical composition comprises at least 0.02% w/w, preferably from about 0.1% to about 2% w/w, of berberine or a biologically equivalent analogue thereof, wherein said amount is based on the total weight of the composition.

In one embodiment, berberine or a biologically equivalent analogue of berberine is the main pharmaceutically acceptable active ingredient.

In one embodiment, berberine or a biologically equivalent analogue of berberine is the only pharmaceutically acceptable active component.

The following examples illustrate some aspects of the invention. The examples are not meant to limit the invention in any way.

Example 1

Mouse skin penetration study of berberine preparation

The following six berberine preparations were compared: c8, 0.125%, 0.3%, G22, G23 and G24.

Preparation

The formulation was as follows:

c8 (cream-based formulation):

aqueous phase

Berberine (0.12%),60 (1%), glycerol (3%), methylparaben (0.1%),

propyl p-hydroxybenzoate (0.02%), NaOH (to adjust pH to 5.5) and EDTA (0.02%).

Oil phase

Stearic acid (7.5%), castor oil (8%), white petrolatum (6%) and SPAN 60 (2%).

0.125% (gel-based formulation):

berberine (0.125%), ethanol (2.5%), glycerol (10%), phenoxyethanol (0.3%), and carbomer.

0.3% (gel-based formulation):

berberine (0.3%), propylene glycol (9.25%), PEG 400 (5.03%), methylparaben (0.1%), propylparaben (0.02%), NaOH (0.4%), EDTA (0.02%), carbomer 934P (1%).

G22 (gel-based formulation):

berberine (0.1%), SDS (0.086%), glycerol (10%),80 (0.5%), methylparaben (0.1%), propylparaben (0.02%), citric acid (0.033%), sodium citrate dihydrate (0.115%), NaOH, EDTA (0.02%), carbomer 934P (0.3%), HEC 250HHX (1.2%). Particle size distribution: 3.83/11.34/27.24 (in the form of D10/D50/D90, where each value refers to a corresponding percentage of particles below the size, i.e., 10% of the particles are less than 3.83, etc.).

G23 (gel-based formulation):

berberine (0.1%), SDS (0.086%), glycerol (10%),80 (0.5%), methylparaben (0.1%), propylparaben (0.02%), citric acid (0.033%), sodium citrate dihydrate (0.115%), NaOH, EDTA (0.02%), carbomer 934P (0.3%), HEC 250HHX (1.2%). Particle size distribution: 1.45/2.85/9.30

G24 (gel-based formulation):

berberine (0.1%), SDS (0)043%), glycerol (10%),80 (0.5%), methylparaben (0.1%), propylparaben (0.02%), citric acid (0.033%), sodium citrate dihydrate (0.115%), NaOH, EDTA (0.02%), carbomer 934P (0.3%), HEC 250HHX (1.2%). Particle size distribution: 1.55/2.86/5.44

The particle size of the G22, G23, and G24 formulations was determined as follows.

Preparing purified water, adding berberine hydrochloride,80 and Sodium Dodecyl Sulfate (SDS). After sufficient dispersion, the mixture is micronized. Thereafter, the particle size was measured by a diffraction analyzer.

Conditions of the experiment

Mice were sacrificed by cervical dislocation. The full thickness flanking skin was removed and placed in a diffusion cell in contact with the receptor, which was 0.01M PBS (pH 7.4 at 37 ℃). The buffer was pumped through the receiver compartment at a flow rate of 3-4 mL/h. 300 μ l of the formulation was added to the skin surface in the donor chamber. Receiver solutions were collected at 0, 1, 2, 3, 4, 6, 8, 10 and 12 hours for HPLC analysis. The skin flux was calculated from the slope of the linear part of the cumulative permeation flux-time curve of berberine hydrochloride.

Results

FIG. 1 shows the cumulative permeation of small chaconine (ng/cm) for all 6 tested formulations²) -a time map. As can be seen, C8 (cream formulation) and G23 (gel-based formulation) penetrated best compared to the other formulations. This is unexpected because, in theory, all six formulations should permeate at similar rates due to the physical properties of berberine in the aqueous phase.

FIG. 2 shows cumulative permeation of small chaconine (ng/cm) of three gel suspension formulations (G22, G23, and G24)²) -a time map. As can be seen, the permeability is positively correlated with the penetration enhancer (SDS) but negatively correlated with berberine size. The permeability of G23 and G24 with the berberine size of D90 less than 10 μm is higher than that of G22 with D90 higher than 10 μm.

Example 2

Mini pig penetration study of berberine formulation

The following berberine preparations were compared: 1) c8, G22, and G23; and 2) 0.125%, 0.30% and G23.

Skin: mini pigs (Lanyu pigs or Lee sung pigs) were skinned up to 700 μm with a resistance >10 kOmega (Millicell-ERS, Millipore).

And (3) penetration experiment:

the pig skin was placed on a diffusion cell with the dermal side in contact with the receptor filled with PBS (pH 7.4 at 37 ℃). To the skin surface in the donor chamber was added 20 μ l of the formulation. After 8 hours, the residual formulation on the skin surface was removed using three dry cotton swabs. At the end of 12 hours and 24 hours of treatment with the formulation, the skin was removed from the diffusion cell and carefully cleaned on the skin surface with three swabs of water. The stratum corneum was removed by tape stripping (tap-striping) 10 times. The skin was then placed on a glass dish and thermally separated in a water bath at 60 ℃ for 90 seconds into epidermis and dermis. The separated epidermis and dermis were weighed and minced with 0.5ml of diluent (1% H)₃PO₄：CH₃OH (1: 1)) extracting. The skin extract was centrifuged at 14,500rpm for 20 min. The berberine hydrochloride concentration in the receiver solution and supernatant from the skin extract was determined by HPLC. Recovery of berberine hydrochloride from the skin was determined by adding a known amount of drug to the skin tissue and treating as described above.

Results

Table 2 summarizes the results of this experiment.

TABLE 2

Run 1 (comparison C8, G22, G23)

Run 2 (compare 0.125%, 0.30%, G23)

The skin permeation result of the mini pig shows that: a) c8 (cream formulation) penetration was surprisingly good; b) the formulation containing berberine granules is released continuously over 24 hours (G22, G23 vs C8, 0.125% and 0.3%); c) g22 and G23 (formulations containing berberine particles) retained more berberine in the epidermis and dermis after 24 hours compared to C8 (cream formulation); d) formulations containing berberine particles penetrated better than formulations containing berberine solution (G23 vs 0.3%); e) even though G23 contained only 0.1% berberine, G23 retained almost the same amount of berberine in the epidermis and more berberine in the dermis after 24 hours compared to the 0.3% formulation; and f) addition of SDS to G23 resulted in increased permeability and local concentration of berberine in the epidermis and dermis compared to other penetration enhancers (ethanol and glycerol in 0.125% formulation and propylene glycol and PEG 400 in 0.3% formulation).

Example 3:

the patient is treated by topical preparation containing berberineLater skin biopsy results

The subjects tested were 56 year old men who received the EGFR inhibitor afatinib for the treatment of non-small cell lung cancer (NSCLC). After receiving afatinib, subjects began applying once daily a topical gel of the G23 formulation on one side of their face and a vehicle gel (G23 without berberine) on the other side.

Bilateral skin biopsies of the subject's nasolabial folds (both sides of the nose) were collected to complete two weeks of topical treatment. Skin specimens were obtained by 1.0cm x 0.5cm incision biopsy and then histologically processed using hematoxylin and eosin (H & E) staining. Evaluation was performed by a trained skin pathologist.

H & E staining results (fig. 3 and 4) show that the hair follicle (follicular) structure of the skin area treated with G23 remained intact and without inflammatory cell infiltration (fig. 3), whereas the hair follicle structure of the skin treated with the vehicle gel was destroyed, with substantial inflammatory cell infiltration in the perifollicular region, with vacuolar changes at the dermal-epidermal junction of the follicular epithelium (fig. 4), indicating the potential anti-inflammatory effect of berberine (G23) for the treatment of skin toxicity associated with EGFR inhibitors.

Example 4:

the patients have facial pathological changes after being treated by the topical preparation containing berberine

During the 4-week treatment period, subjects who started to receive afatinib inhibitor (EGFRI therapy) received G23 on their half sides and vehicle gel (berberine-free G23) on the other half. The subjects were treated as follows 1: 1 to determine the facial side to which study drug was applied. Subjects began using study medication before or within 1 day after the initiation of EGFRI treatment. Study medication was applied once daily (QD) to the designated half side of the face at bedtime (HS).

The number of facial lesions (papules and pustules) was assessed at weekly visits for four weeks. As shown in fig. 5 to 7, the following clear trends were found: the number of lesions on the flank face treated with G23 was significantly lower than on the other half, indicating a potential therapeutic effect of berberine for the treatment of EGFR inhibitors or other targeted therapy-induced or immunotherapy-induced skin toxicity.

Claims

1. A method for the treatment and/or prevention of skin toxicity induced by targeted therapy and/or immunotherapy comprising administering to a patient in need thereof a pharmaceutically effective amount of berberine or a biologically equivalent analogue thereof.

2. The method of claim 1, wherein the targeted therapy is selected from the group consisting of: EGFR inhibitors, Multiple Tyrosine Kinase (MTK) inhibitors, MEK inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, protein kinase b (akt) inhibitors, BRAF inhibitors, HER2 inhibitors, multiple kinase angiogenesis inhibitors, mTOR inhibitors, ALK/c-met inhibitors, multiple kinase Abl inhibitors, BTK inhibitors, HDAC inhibitors, proteasome inhibitors, and RXR agonists.

3. The method of claim 1, wherein the immunotherapy is selected from the group consisting of: cancer vaccines, cytokine agents, cell therapies, immune checkpoint protein inhibitors, and immune checkpoint protein stimulators.

4. The method of claim 1, wherein the skin toxicity is selected from the group consisting of: papulopustular rash, maculopapular rash, erythema, telangiectasias flushing, paronychia and fissures, hair changes, xerosis, mucositis, pruritus and hand-foot skin reactions.

5. The method according to claim 1, wherein the biologically equivalent analogue of berberine is selected from the group consisting of: jateorhizine, palmatine, coptisine, 9-demethylberberine, 9-demethylpalmatine, 13-hydroxyberberine, berrubine, palmatine, 9-O-ethyl berrubine, 9-O-ethyl-13-ethyl berrubine, 13-methyl dihydroberberine N-methyl salt, tetrahydroprotoberberine and its N-methyl salt, and 9-lauryl berberine chloride.

6. The method according to claim 1, wherein the biologically equivalent analogue of berberine is palmatine or coptisine.

7. A method of treating and/or preventing skin toxicity induced by targeted therapy and/or immunotherapy comprising topically applying to the affected skin a pharmaceutically effective amount of a topical pharmaceutical composition comprising berberine or a biologically equivalent analogue thereof.

8. The method of claim 7, wherein the targeted therapy is selected from the group consisting of: EGFR inhibitors, Multiple Tyrosine Kinase (MTK) inhibitors, MEK inhibitors, phosphoinositide 3-kinase (PI3K) inhibitors, protein kinase b (akt) inhibitors, BRAF inhibitors, HER2 inhibitors, multiple kinase angiogenesis inhibitors, mTOR inhibitors, ALK/c-met inhibitors, multiple kinase Abl inhibitors, BTK inhibitors, HDAC inhibitors, proteasome inhibitors, and RXR agonists.

9. The method of claim 7, wherein the immunotherapy is selected from the group consisting of: cancer vaccines, cytokine agents, interferons, interleukin 2, cell therapy, immune checkpoint protein inhibitors, and immune checkpoint protein stimulators.

10. The method of claim 7, wherein the skin toxicity is selected from the group consisting of: papulopustular rash, maculopapular rash, erythema, telangiectasias flushing, paronychia and fissures, hair changes, xerosis, mucositis, pruritus and hand-foot skin reactions.

11. The method according to claim 7, wherein the biologically equivalent analogue of berberine is selected from the group consisting of: jateorhizine, palmatine, coptisine, 9-demethylberberine, 9-demethylpalmatine, 13-hydroxyberberine, berrubine, palmatine, 9-O-ethyl berrubine, 9-O-ethyl-13-ethyl berrubine, 13-methyl dihydroberberine N-methyl salt, tetrahydroprotoberberine and its N-methyl salt, and 9-lauryl berberine chloride.

12. The method according to claim 7, wherein the biologically equivalent analogue of berberine is palmatine or coptisine.

13. The method according to claim 7, wherein the topical pharmaceutical composition comprises at least 0.02% w/w berberine or a biologically equivalent analogue thereof.

14. The method according to claim 7, wherein the topical pharmaceutical composition comprises from about 0.1% to about 2% w/w berberine or a biologically equivalent analogue thereof.

15. The method according to claim 7, wherein berberine or a biologically equivalent analogue thereof is the main pharmaceutically acceptable active component.

16. The method according to claim 7, wherein the berberine or the biologically equivalent analogue thereof is the only pharmaceutically acceptable active component.

17. The method of claim 7, wherein the topical pharmaceutical composition is in the form of a lotion, cream, ointment, paste, gel, spray, suspension, emulsion, foam, patch, powder, and liniment.