WO2021097651A1

WO2021097651A1 - Pharmaceutical composition comprising orlistat and plant-derived lipase inhibitor

Info

Publication number: WO2021097651A1
Application number: PCT/CN2019/119382
Authority: WO
Inventors: 向飞; 杜志博; 彭韪
Original assignee: 中山万汉制药有限公司
Priority date: 2019-11-19
Filing date: 2019-11-19
Publication date: 2021-05-27

Abstract

Disclosed is a pharmaceutical composition comprising orlistat and a plant-derived lipase inhibitor. The plant-derived lipase inhibitor is a polyphenol lipase inhibitor extracted from tea. The results of tests in vitro show that when the molar ratio of orlistat and the polyphenol lipase inhibitor is in the range of 1:10-10:1, the interaction index of inhibiting lipase activity is lower than 1, indicating that the two have a synergistic lipase inhibitory effect. The results of animal testing show that the effect of the composition comprising orlistat and the polyphenol lipase inhibitor on controlling the weight of rats with nutritional obesity is better than that of the group including the independent administration of an orlistat group and the polyphenol lipase inhibitor, further verifying the production of a synergistic effect. In addition, the polyphenol lipase inhibitor can reduce the unpleasant gastrointestinal side effects of orlistat, and alleviate oil spots, fatty/greasy stools and other adverse reactions.

Description

Pharmaceutical composition containing orlistat and plant-derived lipase inhibitor

Technical field

The present invention relates to the technical field of medicine, in particular to a pharmaceutical composition containing orlistat and a plant-derived lipase inhibitor.

Background technique

Orlistat is a lipase inhibitor weight-loss drug developed by Roche Pharmaceuticals under the trade name Xenical. It was the first to be marketed in Europe and the United States in the late 1990s. It was listed in China in 2001 and was approved by China Food in 2005. The Drug Administration approved the conversion to over-the-counter drugs. Its chemical name is N-formyl-L-leucine(s)-1[(2s,3s)3-hexyl-4oxy-2-epoxypropylmethyl]dodecyl ester, also known as tetrahydro Lipostatin (Tetrahydrolipstatin, THL) is a semi-synthetic lipostatin derivative, and its chemical structure is shown in the following formula:

Orlistat forms a covalent bond with the serine residue of gastric pancreatic lipase, which inactivates the enzyme, and cannot hydrolyze triglycerides in food into absorbable fatty acids, thereby reducing the amount of fat absorbed. At present, Orlistat is the only chemical weight loss drug that does not affect appetite and does not act on the central nervous system at home and abroad. It has superior safety characteristics. An article published by Bray GA in The Lancet is called Management of Obesity's article described orlistat as the "safest" weight loss drug. However, orlistat can only inhibit 30% of fat absorption after oral administration. Bray GA et al. published an article in "The Lancet" entitled Management of Obesity rated the weight loss effect of orlistat as " Modest (moderate)", not as good as the recently marketed appetite suppressant type of weight-loss drugs. In view of the superior safety of orlistat, it is necessary to improve the weight loss effect of orlistat preparations through pharmacological or pharmacological means, so as to expand its clinical application to deal with the increasingly severe obesity situation.

The only lipase inhibitors that have been approved for marketing at home and abroad are orlistat and cetilistat. The former can be obtained by total synthesis or semi-synthesis, and the latter can only be obtained by total synthesis. In addition to chemical compounds, plants also contain a wealth of lipase inhibitors, and their structure types include polyphenols, saponins, terpenes, flavonoids, alkaloids, etc. The lipase inhibitor extracted from tea not only has the advantages of wide sources, diverse structures, high specificity, low toxicity, etc., but also because it acts on different active sites of lipase, it may produce synergistic weight loss with orlistat effect.

Nakai M et al. (Journal of Agricultural and Food Chemistry, 2005,53(11):4593～4598) measured the lipase inhibitory activity of 54 polyphenolic compounds extracted from tea, and found that related compounds inhibit fat The IC50 of the enzyme is between 0.048～>20μM, which is generally lower than that of other plant-derived lipase inhibitors described by Li Qiang et al. in their review (Natural Products Research and Development. 2015, 27: 360～366) However, in the prior art, there is no teaching on the specific binding sites of lipase inhibitors extracted from tea and lipase, and there is no teaching on whether orlistat and lipase inhibitors extracted from tea produce synergistic effects after being combined.

Summary of the invention

In order to overcome the technical problems in the prior art, the purpose of the present invention is to provide a pharmaceutical composition containing orlistat and polyphenol lipase inhibitors extracted from tea, the active ingredients in the pharmaceutical composition It can produce a synergistic lipase inhibitory effect, and has a good weight loss effect, and can be used to treat and prevent obesity.

The present invention achieves the above objectives through the following technical solutions:

The present invention provides a pharmaceutical composition for the treatment and prevention of obesity. The pharmaceutical composition comprises orlistat and a plant-derived lipase inhibitor. Specifically, the plant-derived lipase inhibitor is from tea Polyphenolic lipase inhibitor extracted from it.

Specifically, the lipase inhibitor extracted from the tea leaves is selected from at least one compound whose structural formula is shown in the following formula.

The in vitro test results show that the molar ratio (ratio of the amount of substance) of orlistat in the pharmaceutical composition for the treatment and prevention of obesity provided by the present invention and the polyphenol lipase inhibitor extracted from tea is 1: In the range of 10-10:1, the interaction index of inhibiting lipase activity is lower than 1, indicating that the two have a synergistic lipase inhibitory effect.

Preferably, when the polyphenol lipase inhibitor is a compound represented by formula I (compound 1), the molar ratio of compound 1 to orlistat is 1:9 to 1:6. More preferably, the molar ratio of compound 1 to orlistat is 1:7.75.

Preferably, when the polyphenol lipase inhibitor is a compound represented by formula II (compound 2), the molar ratio of compound 2 to orlistat is 1:4 to 1:2. More preferably, the molar ratio of compound 2 to orlistat is 1:3.

Preferably, when the polyphenol lipase inhibitor is a compound represented by formula III (compound 3), the molar ratio of compound 3 to orlistat is 1:2 to 2:1. More preferably, the molar ratio of compound 3 to orlistat is 1.25:1.

Preferably, when the polyphenol lipase inhibitor is a compound represented by formula VI (compound 4), the molar ratio of compound 4 to orlistat is 1:4 to 1:1. More preferably, the molar ratio of compound 4 to orlistat is 1:2.5.

On the other hand, animal test results show that orlistat and polyphenol lipase inhibitors extracted from tea in the pharmaceutical composition for the treatment and prevention of obesity provided by the present invention are the most preferred according to in vitro tests. The molar ratio of the combination, calculated by mole ratio, the total dose is the same, and the doses of orlistat and polyphenol lipase inhibitor in the composition are both lower than the two separate administration groups under the premise that the composition has an effect on nutrition The weight control effect of obese rats is better than that of the orlistat group and the polyphenol lipase inhibitor alone administration group, further verifying the synergy.

At the same time, the present inventors were surprised to find that when orlistat was used in combination with polyphenol lipase inhibitors, the number of animals with grease distribution in rat hair in the test group was significantly reduced, suggesting the drug combination provided by the present invention The compound can reduce the probability of the appearance of grease in rat hair, and the polyphenol lipase inhibitor can reduce the unpleasant gastrointestinal side effects of orlistat, and alleviate adverse reactions such as oil spots, fat/oily stools and the like.

The pharmaceutical composition for the treatment and prevention of obesity provided by the present invention can be used directly, or one or more pharmaceutical excipients can be added and an oral preparation, such as a powder, can be prepared by a preparation method well known to those skilled in the art. , Granules, capsules, tablets, pills, etc. The preparation methods well-known to those skilled in the art can refer to but not limited to the editor-in-chief Cui Fude, "Pharmaceuticals" (7th edition) (published by People's Medical Publishing House), which is incorporated herein by reference. Specifically, orlistat, the polyphenol lipase inhibitor extracted from the tea can be mixed with at least one pharmaceutically acceptable excipient, such as citric acid or dicalcium phosphate , Or: (a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, such as cellulose derivatives, starch, alginate, gelatin, Polyvinylpyrrolidone, sucrose and acacia gum; (c) disintegrants, such as agar, calcium carbonate, potato or tapioca starch, alginic acid, croscarmellose sodium, complex silicate and sodium carbonate; d) Solution retarders, such as paraffin; (e) absorption enhancers, such as quaternary ammonium compounds; (f) wetting agents, such as cetyl alcohol and glyceryl monostearate, magnesium stearate, etc.; (g ) Adsorbents, such as kaolin and bentonite; and (h) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyglycol, sodium lauryl sulfate, or mixtures thereof.

Preferably, the pharmaceutical composition provided by the present invention is made into a capsule by adding an appropriate amount of pharmaceutical excipients.

Compared with the prior art, the advantages of the present invention are:

The present invention discloses for the first time that the composition of orlistat and polyphenol lipase inhibitors (compounds represented by formula I-VI) extracted from tea has a synergistic lipase inhibitory effect, and it is applied to the treatment or prevention of obesity The disease has a good synergistic gain effect, and the weight control effect on the nutritionally obese rats is significantly better than the orlistat group and the polyphenol lipase inhibitor alone administration group. At the same time, surprisingly, the polyphenols Lipase inhibitors can also reduce the unpleasant gastrointestinal side effects of orlistat, relieve oily spots, fat/oily stools and other adverse reactions, achieving unexpected results.

Description of the drawings

Figure 1 is a graph showing the relationship between the interaction index (Y) of compound 1 and orlistat and the molar ratio of the two (molar ratio range 1:10-10:1).

Figure 2 is a graph showing the relationship between the interaction index (Y) and the molar ratio of the compound 1 and orlistat (preferred molar ratio range).

Figure 3 is a diagram showing the relationship between the interaction index (Y) of compound 2 and orlistat and the molar ratio of the two (molar ratio range 1:10-10:1).

Figure 4 is a graph showing the relationship between the interaction index (Y) and the molar ratio of the compound 2 and orlistat (preferred molar ratio range).

Figure 5 is a diagram showing the relationship between the interaction index (Y) of compound 3 and orlistat and the molar ratio of the two (molar ratio range 1:10-10:1).

Fig. 6 is a graph showing the relationship between the molar ratio of the interaction index (Y) of compound 3 and orlistat (preferred range of molar ratio).

Figure 7 is a diagram showing the relationship between the molar ratio of the interaction index (Y) of compound 4 and orlistat (molar ratio range 1:10-10:1).

Figure 8 is a graph showing the relationship between the molar ratio of the interaction index (Y) of compound 4 and orlistat (preferred range of molar ratio).

Detailed ways

The present invention will be further described in detail below with reference to the drawings and specific embodiments of the specification. The embodiments are only used to explain the present invention and not used to limit the scope of the present invention. The test methods used in the following examples are conventional methods unless otherwise specified; the materials and reagents used, unless otherwise specified, are commercially available reagents and materials.

First, the applicant entrusted Zhongshan Wanyuan New Drug Research and Development Co., Ltd. (hereinafter referred to as "Zhongshan Wanyuan") to synthesize the polyphenol lipase inhibitors in the pharmaceutical composition of the present invention, according to the quality report provided by Zhongshan Wanyuan It is shown that the purity of the compounds 1-4 are all above 99.7%.

Secondly, the present invention adopts the lipase inhibitory activity assay method disclosed by Nakai M et al. (Journal of Agricultural and Food Chemistry, 2005, 53(11):4593～4598), and uses the interaction index (Y) as an indicator to investigate The synergistic inhibitory effect of orlistat and a single polyphenol lipase inhibitor combined with a molar ratio of 1:10 to 10:1 on lipase activity, and the optimal molar ratio was screened. The results are shown in Figure 1 -8 shown. Here, the calculation method of the interaction index (Y) is shown in the following formula.

In the above formula, IC50(A) and IC50(B) respectively represent the IC50 value of lipase when orlistat is treated with one of the corresponding compounds 1-4 separately, and IC50(mixA) and IC50(mixB) respectively represent mixed The concentration of orlistat and the corresponding polyphenol lipase inhibitor when the system produces half inhibition of lipase.

It can be seen from Figures 1, 3, 5, and 7 that orlistat combined with compound 1-4, respectively, has a low interaction index (Y) for inhibiting lipase activity in the molar ratio range of 1:10～10:1. In 1, it has a synergistic lipase inhibitory effect. And when compound 1 is combined with orlistat, the effect of inhibiting lipase activity is better in the molar ratio range of 1:9 to 1:6; when compound 2 is combined with orlistat, the effect is between 1:4 to 1: The effect of inhibiting lipase activity is better in the range of molar ratio of 2; when compound 3 is combined with orlistat, the effect of inhibiting lipase activity is better in the range of molar ratio of 1:2 to 2:1; when compound 4 In combination with orlistat, the effect of inhibiting lipase activity is better in the molar ratio range of 1:4 to 1:1.

It can be seen from Figure 2 that when compound 1 is combined with orlistat, the lipase activity inhibition effect is best at a molar ratio of 1:7.75.

It can be seen from Figure 4 that when compound 2 is combined with orlistat, the lipase activity inhibition effect is the best at a molar ratio of 1:3.

It can be seen from Figure 6 that when compound 3 is combined with orlistat, the lipase activity inhibition effect is the best at a molar ratio of 1.25:1.

It can be seen from Fig. 8 that when compound 4 is combined with orlistat, the lipase activity inhibition effect is the best at a molar ratio of 1:2.5.

Test Example 1. The weight loss effect of the combination of orlistat and polyphenol lipase inhibitors on nutritionally obese rats

This test method adopts the method disclosed in the "Slimming Effect of Orlistat on Nutritionally Obese Rats" published by Liu Jingru et al. (Chinese Journal of Experimental Pharmacology, 2013,19(07):186-188.). The following administration groups:

1) Blank control group;

2) Model group;

3) Orlistat group;

4) Test group A1-4;

5) Test group B1-4,

6) Test group C,

7) Test group D, 15 rats in each group.

Among them, the definitions and treatment methods of the blank control group, model group and orlistat group are consistent with those disclosed by Liu Jingru et al.; the experimental group A1-4 was fed with nutrient feed while the rats were fed with compounds 1-4 respectively. The pretreatment method before gavage is the same as the orlistat group, and the dosage is the same as that of the orlistat group, which is 0.121mmol/kg/d. In the test group B1-4, while feeding the rats with nutritious feed, the compound 1-4 and orlistat were combined according to the most preferred substance ratio measured in the in vitro test, and then gavage the rats before gavage. The pretreatment method is the same as that of the orlistat group, and the total dose of the two active ingredients in the combination is the same as that of the orlistat group, which is 0.121mmol/kg/d. In experimental group C, while feeding the rats with nutritive feed, they were given epigallocatechin gallate (EGCG, purchased from Shaanxi Sovere Natural Products Co., Ltd.) by gavage. The pretreatment method before gavage was the same as that of Orly In the stat group, the dose is the same as the orlistat group in terms of the amount of substance, that is, 0.121mmol/kg/d. In experimental group D, while feeding the rats with nutritious feed, they were given a combination of epigallocatechin gallate and orlistat at a ratio of 1:1 and then gavage. The pretreatment method before gavage was the same In the orlistat group, the total dose of the two active ingredients in the combination is the same as the orlistat group, which is 0.121mmol/kg/d. After 7 weeks of continuous administration, the weight changes of the animals in each group were observed, and the number of animals with fat distribution in the hair of nutritionally obese rats was counted, and the probability of fat was calculated. The results are shown in Tables 1 and 2 below.

Table 1 Results of changes in body weight of rats in each test group after 7 weeks of administration

Note: a P< 0.01 compared with the blank control group, t test

b Compared with the model group, P< 0.01, t test

b*Compared with the model group, P< 0.01, but compared with the orlistat group, P> 0.05, both t test

c and Orlistat group ratio P< 0.01, t test

d Compared with the corresponding group A, P< 0.01, t test

The results showed that when compounds 1 to 4 were administered alone, their inhibitory effects on weight gain in nutritionally obese rats were equivalent to orlistat, but they were administered in combination with orlistat, and the total dose (depending on the substance) When the meter) is equivalent to orlistat administered alone, the combined administration has a significantly better effect on the weight gain of nutritionally obese rats than that of orlistat administered alone (P< 0.01), indicating that it produces Synergistic weight loss effect. In addition, the present invention confirms that the combined application of EGCG and orlistat has a slightly better effect on inhibiting weight gain in rats than the EGCG group alone or the orlistat group alone, but there is no significant difference, indicating that EGCG and orlistat His combination did not produce a synergistic effect. The above results indicate that only when a specific polyphenol lipase inhibitor is combined with orlistat can a synergistic weight loss effect be obtained.

Table 2 Results of changes in body weight of rats in each test group after 7 weeks of administration

The results showed that after orlistat was administered alone for 1 week, the number of animals with grease distribution in rat hair was 9 and 15 after 7 weeks, while the combination of compound 1-4 and orlistat at the same time could significantly reduce The number of animals with grease distribution in rat hair, and only 2 rats had hair oil spills 7 weeks after administration, indicating that the combination of compound 1-4 of the present invention and orlistat can reduce orlistat-induced hair in rats The probability of occurrence of grease, the compounds 1-4 can reduce the unpleasant gastrointestinal side effects of orlistat, and alleviate adverse reactions such as oil spots, fat/oily stools and the like. In addition, the present invention confirms that the combination of EGCG and orlistat does not significantly reduce the probability of fat in rat hair. The above results show that only when a specific polyphenol lipase inhibitor is combined with orlistat can it be used. Obtain a synergistic effect of inhibiting hair oil spills.

Example 1 Capsule preparation

Example 1 Preparation of Capsules Containing Orlistat and Compound 1

Prescription: (According to 1000 tablets)

Preparation Process:

Pass sodium lauryl sulfate, crospovidone, sodium starch glycolate, microcrystalline cellulose, orlistat, and compound 1 powder through an 80-mesh sieve and set aside. Weigh the prescription amount of sodium lauryl sulfate, crospovidone, sodium starch glycolate, microcrystalline cellulose, orlistat, compound 1 powder, and first add sodium lauryl sulfate and crospovidone The ketone and sodium starch glycolate are mixed uniformly, then microcrystalline cellulose, orlistat, and compound 1 are added and mixed uniformly, and passed through an 80-mesh sieve twice. The mixed powder is slowly added to a 50% ethanol solution containing 10% povidone K30 to make soft material, 20 mesh squeezed through a sieve to make granules, and the wet granules are dried in a blast drying oven at 30°C for 6 hours, and the 20 mesh sieve is taken out Take the whole capsule and put it into No. 0 capsule to get it.

Example 2 Tablet preparation

Example 1 Preparation of tablets containing orlistat and compound 2

prescription:

Preparation Process:

Weigh orlistat and compound 2 according to the prescription, use microcrystalline cellulose as filler, croscarmellose sodium and polyvinylpyrrolidone as disintegrants, and 5% PVP 60% ethanol solution as binder. Micronized silica gel is a glidant, which is granulated in a fluidized bed in one step, and then compressed into tablets.

Example 3 Preparation of granules

Example 3 Preparation of granules containing orlistat and compound 3

prescription:

Preparation Process:

Weigh the prescription amount of orlistat, compound 3, starch, dextrin, and sucrose powder and mix them evenly. In addition, an appropriate amount of 80% ethanol is added to the mixed powder and mixed evenly to form a soft material, which is made into wet granules through an 18-mesh nylon sieve, dried at about 60°C, and sized by a 20-mesh sieve.

Example 4 Preparation of sustained-release tablets

Example 4 Preparation of sustained-release tablets containing orlistat and compound 4

prescription:

Preparation process: According to the preparation process of the sustained-release tablet, it can be prepared into a sustained-release tablet.

The above are only the preferred embodiments of the present invention. It should be pointed out that the above preferred embodiments should not be regarded as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. For those of ordinary skill in the art, without departing from the spirit and scope of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

A pharmaceutical composition for the treatment and prevention of obesity, which is characterized in that it contains orlistat and a plant-derived lipase inhibitor, and the molar ratio of the orlistat to the plant-derived lipase inhibitor is 1:10～10:1.
The pharmaceutical composition according to claim 1, wherein the plant-derived lipase inhibitor is selected from at least one of the compounds represented by formula I, formula II, formula III, and formula VI;
The pharmaceutical composition according to claim 2, wherein the plant-derived lipase inhibitor is selected from the group consisting of compounds of formula I, the compound of formula I and orlistat The molar ratio is 1:9 to 1:6.
The pharmaceutical composition according to claim 3, wherein the molar ratio of the compound represented by formula I to orlistat is 1:7.75.
The pharmaceutical composition according to claim 2, wherein the plant-derived lipase inhibitor is selected from the group consisting of compounds represented by formula II, the compounds represented by formula II and orlistat The molar ratio is 1:4 to 1:2.
The pharmaceutical composition according to claim 5, wherein the molar ratio of the compound represented by formula II to orlistat is 1:3.
The pharmaceutical composition according to claim 2, wherein the plant-derived lipase inhibitor is selected from the group consisting of compounds represented by formula III, the compounds represented by formula III and orlistat The molar ratio is 1:2 to 2:1.
The pharmaceutical composition according to claim 7, wherein the molar ratio of the compound represented by formula III to orlistat is 1.25:1.
The pharmaceutical composition according to claim 2, wherein the plant-derived lipase inhibitor is selected from the group consisting of compounds represented by formula VI, the compounds represented by formula VI and orlistat The molar ratio is 1:4 to 1:1.
The pharmaceutical composition according to claim 9, wherein the molar ratio of the compound represented by formula VI to orlistat is 1:2.5.