CN108774264A - Phosphocholine analogs, Preparation method and use - Google Patents

Abstract

The invention provides a lecithin analog represented by formula (1), and the invention also provides a preparation method of the lecithin analog and its use in the preparation of antitumor drugs.

Description

Lecithin analogue, preparation method and use

technical field

The invention relates to the fields of medicinal chemistry and therapeutics, in particular to a lecithin analogue, a preparation method and its use as an antitumor drug.

Background technique

Lecithin (that is, phosphatidylcholine) is one of the main components of phospholipids. Lecithin quickly transmits the instructions of the brain. The faster the information transmission speed, the stronger the memory. It is a brain food. It can enhance baby's intelligence and improve learning efficiency. Especially the fetus has a great demand for lecithin during growth and development. In addition, lecithin can also prevent fatty liver and alcoholic liver, effectively protecting liver health. The amphiphilic nature of lecithin makes it emulsifying, which can fully mix the water and oil in the body, avoid a large amount of water loss, and prevent rough and aging skin; it can turn excess fat into smaller milk droplets and excrete it to help puerpera or Obese people get back into shape as soon as possible. Lecithin can also convert cholesterol and fatty acid in the blood into very fine particles, which are discharged from the blood vessels, thereby increasing the content of high-density lipoprotein, restoring the elasticity of the blood vessels, and smoothing the blood flow. It is known as the "vascular scavenger".

Cancer chemotherapy refers to the use of drugs to treat cancer, which can eliminate cancer cells that have spread to various parts of the body. However, chemotherapy is often accompanied by serious side effects. Self-assembled nano-preparations can effectively improve the efficacy of chemotherapy drugs and reduce toxicity. Lecithin is an amphiphilic molecule consisting of a hydrophilic head and a hydrophobic tail that can self-assemble into nanoparticles. Through chemical bonding, the monofatty acylglycerol lecithin is combined with the compound molecule with anticancer effect to form a chemotherapeutic drug-lecithin complex. The complex can not only prepare a corresponding liposome drug delivery system through self-assembly, but also effectively reduce drug toxicity.

Camptothecin (CPT) is an alkaloid extracted from the unique plant camptothecin in my country. Camptothecin and its derivatives have formed an important class of anticancer drugs. The closed α-hydroxylactone ring in the E ring structure of camptothecin is an essential structure for maintaining anti-tumor activity, but this α-hydroxylactone ring is easily hydrolyzed in the human body to form a carboxylate structure. The form is easy to combine with human serum albumin and make it lose anti-tumor activity. What's more serious is that the sodium salt in the open-loop form has great toxic and side effects on the urinary system and digestive system after being metabolized by the kidneys. In addition, most active derivatives of camptothecin are poorly soluble in water, which seriously hinders the popularization and application of camptothecin derivatives in clinical practice.

There are many studies on the preparation methods of camptothecin derivatives, and some literatures are given below for reference.

US Patent 104894, No. 1990-3-1

PCT patent application 2001009139, No. 2001-2-8

PCT patent application 9602546, No. 1996-7-12

J Biol Chem, 1985, 260, 14873-14878

J Med Chem, 1991, 34(1), 98-107

J Med Chem, 1998, 41(1), 31-37

Bioorg Med Chem Lett, 2002, 12(9), 1241-1244

Bioorg Med Chem Lett, 2003, 13(21), 3739-3741

Bioorg Med Chem, 2004, 12(15), 4003-4008

Bioorg Med Chem, 2004, 12(13), 3657-3662

Chem Pharm Bull, 1991, 39, 3183-3188

Cancer Res, 1993, 53, 1577-1582

Cancer Res, 1995, 55, 753-760

N Y Acada Sci, 1996, 803, 231-246

Arch Pharm Res, 1998, 21, 581-590

J Med Chem, 1995, 38(3), 395-401

Contents of the invention

One object of the present invention is to provide a class of lecithin analogs combined with anticancer drugs, which is beneficial to improve the anticancer activity and bioavailability of the drugs.

Another object of the present invention is to provide a preparation method of such lecithin analogues.

Another object of the present invention is to provide the application of the lecithin analogs as antitumor drugs.

In order to achieve the above object, the present invention provides a lecithin analog represented by formula (1).

Wherein, R is selected from:

-CH ₂ CH ₂ -SS-CH ₂ CH ₂ -, -CH ₂ -SS-CH ₂ -,

R1 is selected from: -OCOCH ₂ (CH ₂ ) ₁₅ CH ₃ , -OCOCH ₂ (CH ₂ ) ₆ CH=CH(CH ₂ ) ₇ CH ₃ , -OCOCH ₂ (CH ₂ ) ₁₃ CH ₃ ;

R2 is selected from: -H, -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ OCH ₃ , -OCH ₂ CH ₂ OCH ₃ .

The present invention provides a method for preparing a lecithin analog shown in (1), comprising the steps of:

(1) In a dry organic solvent, camptothecin or its derivatives and the corresponding carboxylic acid carry out an esterification reaction under the action of a coupling agent and a catalyst to obtain an intermediate a with a structure of formula (2);

Wherein, R is selected from:

-CH ₂ -CH ₂ -SS-CH ₂ -CH ₂ -, -CH ₂ -SS-CH ₂ -,

R2 is selected from: -H, -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ OCH ₃ , -OCH ₂ CH ₂ OCH ₃ ;

(2) Intermediate a reacts with the corresponding 1-fatty acyl-Sn-glycerol-3-phosphocholine under the action of a coupling agent and a catalyst to prepare the corresponding lecithin analog; the carboxylic acid Selected from 3,3'-dithiodipropionic acid, dithioglycolic acid, cystine; the 1-fatty acyl-Sn-glycerol-3-phosphocholine is selected from 1-stearyl Acyl-Sn-glycerol-3-phosphocholine, 1-oleoyl-Sn-glycerol-3-phosphocholine, 1-palmitoyl-Sn-glycerol-3-phosphocholine.

Wherein, the organic solvent described in the method for preparing lecithin analogs is methylene chloride, dimethyl sulfoxide and N, N-dimethylformamide; the coupling agent is N, N-dicyclohexyl Carbodiimide (DCC), N, N-carbonyldiimidazole (CDI) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCl); the The catalysts are pyridine and 4-dimethylaminopyridine (DMAP).

The present invention also relates to various formulations comprising the compound of the present invention as an active ingredient together with a pharmacodynamically acceptable carrier.

"Pharmacologically acceptable carrier" refers to: one or more compatible solid or liquid filler or gel substances, which are suitable for human use, and have sufficient purity and low toxicity. "Compatibility" herein means that the components of the composition can be blended with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include sugars (such as glucose, sucrose, lactose, etc.), starches (such as corn starch, potato starch, etc.), cellulose and its derivatives (such as sodium carboxymethylcellulose, ethyl Sodium cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybeans, sesame oil, peanut oil, olive oil, etc.), multivariate Alcohol (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifier (such as Tween), lubricant (such as sodium lauryl sulfate), coloring agent, flavoring agent, stabilizer, antioxidant, preservative, No pyrogen water etc.

The present invention also relates to the application of the compound described in the present invention in the preparation of antitumor drugs.

The in vitro activity screening experiment shows that the lecithin analog has obvious anti-tumor effect and good dose-dependent relationship. Using the human ovarian cancer cell line HXB1309H as the test cell line, the half maximal inhibitory concentration (IC50) of lecithin analogues was determined, and the experimental results of some samples are shown in Table 1:

Table 1: The half maximal inhibitory concentration of lecithin analogs to XB1309

Detailed ways

Below in conjunction with embodiment, the present invention is described in further detail:

It has a lecithin analog represented by formula (1).

Wherein, R is selected from:

-CH ₂ -CH ₂ -SS-CH ₂ -CH ₂ -, -CH ₂ -SS-CH ₂ -,

R1 is selected from: -OCOCH ₂ (CH ₂ ) ₁₅ CH ₃ , -OCOCH ₂ (CH ₂ ) ₆ CH=CH(CH ₂ ) ₇ CH ₃ , -OCOCH ₂ (CH ₂ ) ₁₃ CH ₃ ;

R2 is selected from: -H, -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ OCH ₃ , -OCH ₂ CH ₂ OCH ₃ .

The present invention provides a method for preparing a lecithin analog shown in (1), comprising the steps of:

(1) In a dry organic solvent, camptothecin or its derivatives and the corresponding carboxylic acid are subjected to an esterification reaction under the action of a coupling agent and a catalyst to obtain an intermediate a having a structure of formula (2);

Wherein, R is selected from:

-CH ₂ -CH ₂ -SS-CH ₂ -CH ₂ -, -CH ₂ -SS-CH ₂ -

R2 is selected from: -H, -OH, -OCH ₃ , -OCH ₂ CH ₃ , -OCH ₂ CH ₂ OCH ₃ , -OCH ₂ CH ₂ OCH ₃ ;

(2) Intermediate a reacts with the corresponding 1-fatty acyl-Sn-glycerol-3-phosphocholine under the action of a coupling agent and a catalyst to prepare the corresponding lecithin analog; the carboxylic acid Selected from 3,3'-dithiodipropionic acid, dithioglycolic acid, cystine; the 1-fatty acyl-Sn-glycerol-3-phosphocholine is selected from 1-stearyl Acyl-Sn-glycerol-3-phosphocholine, 1-oleoyl-Sn-glycerol-3-phosphocholine, 1-palmitoyl-Sn-glycerol-3-phosphocholine.

Wherein, the organic solvent described in the method for preparing lecithin analogs is methylene chloride, dimethyl sulfoxide and N, N-dimethylformamide; the coupling agent is N, N-dicyclohexyl Carbodiimide (DCC), N, N-carbonyldiimidazole (CDI) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC.HCl); the The catalysts are pyridine and 4-dimethylaminopyridine (DMAP).

The present invention also relates to various formulations comprising the compound of the present invention as an active ingredient together with a pharmacodynamically acceptable carrier.

"Pharmacologically acceptable carrier" refers to: one or more compatible solid or liquid filler or gel substances, which are suitable for human use, and have sufficient purity and low toxicity. "Compatibility" herein means that the components of the composition can be blended with the compound of the present invention and with each other without significantly reducing the efficacy of the compound. Some examples of pharmaceutically acceptable carriers include sugars (such as glucose, sucrose, lactose, etc.), starches (such as corn starch, potato starch, etc.), cellulose and its derivatives (such as sodium carboxymethylcellulose, ethyl Sodium cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil

etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween), lubricants (such as sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants , preservatives, pyrogen-free water, etc.

The present invention also relates to the application of the compound described in the present invention in the preparation of antitumor drugs.

The lecithin analogs and preparation methods of the present invention are described in more detail in the following examples, but the examples do not constitute a limitation to the present invention.

Example 1 Preparation of camptothecin-20-O-(3,3'-dithiodipropionyl) monoester

Dissolve 2.1 g (0.01 mol) of 3,3'-dithiodipropionic acid in 300 ml of dichloromethane, add 1 g of dimethylaminopyridine, 3.5 g (0.01 mol) of camptothecin, and 3 g of N, N'-dicyclohexylcarbimide, stirred at room temperature for 24 hours, filtered to remove the precipitate, the filtrate was concentrated under reduced pressure to remove the solvent, and the concentrated residue was subjected to silica gel column chromatography (dichloromethane:methanol=20:1), 3.35 g of light yellow camptothecin-20-O-(3,3'-dithiodipropionyl) monoester was obtained, with a yield of 62%.

1HNMR (300MHz, DMSO-d6, ppm): δ0.96 (3H, t, H-19), 2.10 (2H, m, H _- 18), 2.58-2.72 (4H, m, _-OOCC H2CH2S -), 2.87-2.99 (4H, m, -OOCCH ₂ CH ₂ S-), 5.11 (2H, s, H-5), 5.50 (2H, s, H-17), 6.89 (1H, d, NH ), 7.14(H, s, H-14), 7.72(1H, q, H-10), 7.87(H, t, H-11), 8.00(1H, d, H-12), 8.14(1H, d, H-9), 8.42 (1H, t, NH), 8.65 (1H, s, H-7).

ESIMS m/z: 539.3(M-1) ^-

Example 2 Preparation of 1-stearyl-2-(camptothecin-20-O-(3,3'-dithiodipropionyl) monoester) glycerol-3-phosphocholine (lecithin similar thing)

Dissolve 0.540 g (0.001 mol) of camptothecin-20-O-(3,3'-dithiodipropionyl) monoester in 50 ml of dimethyl sulfoxide, add 0.5 g of 1-ethyl-( 3-Dimethylaminopropyl) carbodiimide hydrochloride, 0.1 g of 4-dimethylaminopyridine, at room temperature, after stirring for 3 hours, add 0.523 g (0.001 mole) of 1-stearyl-Sn-glycerine Alcohol-3-phosphocholine, at room temperature, stirred for 72 hours, poured the reaction solution into 500 ml of ice water, filtered, and dried the filter cake in vacuum at 50°C, and the filter cake was subjected to silica gel column chromatography (methanol: chloroform: Water=25:65:2), to get yellow 1-stearyl-2-(camptothecin-20-O-(3,3'-dithiodipropionyl)monoester)glycerol-3- Phosphorylcholine 0.21 g (ie lecithin analogue), yield 20%.

1HNMR (300MHz, DMSO-d6, ppm): 2.58-2.72 (4H, m, -OOCC H ₂ CH ₂ S-), 2.87-2.99 (4H, m, -OOCCH ₂ CH ₂ S-), 3.23 (12H ,m,-NCH3,-CH2-N-),3.56-4.17(4H m,-OC ₂ H ₄ N-),5.13(2H,s,H-5),5.55(2H,s,H-17) , 6.89 (1H, d, NH), 7.14 (H, s, H-14), 7.72 (1H, q, H-10), 7.87 (H, t, H-11), 8.00 (1H, d, H -12), 8.14 (1H, d, H-9), 8.42 (1H, t, NH), 8.65 (1H, s, H-7).

Example 3 prepares lecithin analog self-assembled liposomes

Dissolve 0.05 g of 1-stearoyl-2-(camptothecin-20-O-(3,3'-dithiodipropionyl)monoester)glycerol-3-phosphocholine in 5 mL of methanol , under the action of ultrasound, add 25 ml of distilled water at one time, after 10 minutes of ultrasound, quickly blow off methanol, and freeze-dry the residue to obtain nano-scale particles (particle size ≤ 200nm), which is the self-assembled lipid containing lecithin analogues plastid.

Claims (6)

1. phosphocholine analogs shown in formula (1)

Wherein, R_-It is selected from：

-CH₂-CH₂-S-S-CH₂-CH₂-、-CH₂-S-S-CH₂-、

R_-1-It is selected from：-OCOCH₂(CH₂)₁₅CH₃、-OCOCH₂(CH₂)₆CH=CH (CH₂)₇CH₃、-OCOCH₂(CH₂)₁₃CH₃；

R_-2-It is selected from：-H,-OH,-OCH₃、-OCH₂CH₃、-OCH₂CH₂OCH₃、-OCH₂CH₂OCH₃。

2. the method for preparing phosphocholine analogs described in claim 1, includes the following steps：

(1) in dry organic solvent, the effect of camptothecine or derivatives thereof and corresponding carboxylic acid in coupling agent and catalyst Lower carry out esterification, obtains the intermediate a with formula (2) structure；

Wherein, R_-It is selected from：

-CH₂-CH₂-S-S-CH₂-CH₂-、-CH₂-S-S-CH₂-、

R_-2-It is selected from：-H,-OH,-OCH₃、-OCH₂CH₃、-OCH₂CH₂OCH₃、-OCH₂CH₂OCH₃；

(2) intermediate a is anti-with corresponding 1- fatty acyls-Sn- glycerine -3- phosphocholines under the action of coupling agent and catalyst It answers, corresponding phosphocholine analogs is made；The carboxylic acid is selected from 3,3'- dithiodipropionic acids, dithioglycollic acid, Guang ammonia Acid；1- fatty acyl-Sn- glycerine -3- the phosphocholines are selected from 1- stearoyl-Sn- glycerine -3- phosphocholines, 1- oil Acyl-Sn- glycerine -3- phosphocholines, 1- palmityl-Sn- glycerine -3- phosphocholines.

3. the method for preparing phosphocholine analogs according to claim 2, it is characterised in that the organic solvent is two Chloromethanes, dimethyl sulfoxide (DMSO) and n,N-Dimethylformamide；The coupling agent is N, N- dicyclohexylcarbodiimides (DCC), N, N- carbonyl dimidazoles (CDI) and 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides (EDC.HCl)；Described Catalyst is pyridine and 4-dimethylaminopyridine (DMAP).

4. a kind of pharmaceutical composition contains acceptable carrier in any compound described in claim 1 and pharmacodynamics.

5. pharmaceutical composition according to claim 4, it is characterised in that the pharmaceutical composition can be tablet, capsule, ball Agent, injection, sustained release preparation.

6. phosphocholine analogs application in preparation of anti-tumor drugs described in claim 1.