Composition of alfanotide and semeglutide and application of composition in preparation of hypolipidemic drugs

Abstract

The invention relates to the technical field of blood fat reduction, in particular to a composition of alfanotide and semeglutide and application of the composition in preparation of blood fat reducing drugs. After the alfanopeptide and the semeglutide are combined for use, the synergistic blood fat reducing effect is realized, the content of TC and TG in serum is remarkably reduced, a new direction is provided for research and development of blood fat reducing medicines, and meanwhile, the application field of the alfanopeptide is widened.

Description

Technical Field

[0001] The present invention relates to the technical field of lowering blood lipids, and in particular to a combination of afamelanotide and semaglutide and an application thereof in preparing a blood lipid-lowering drug. Background Art

[0002] Cardiovascular disease is one of the main causes of serious harm to human health. Hyperlipidemia, when plasma lipid concentrations exceed the normal range, is a significant risk factor for the development of cardiovascular diseases such as coronary heart disease and myocardial infarction. Its most direct hazard is the development of atherosclerosis. Therefore, research on lipid-lowering drugs is crucial.

[0003] Traditional lipid-lowering drugs include statins, fibrates, niacin, ezetimibe, and bile acid sequestrants. Atorvastatin and rosuvastatin have become blockbuster drugs in the global pharmaceutical market. Furthermore, fibrates (represented by gemfibrozil and ciprofibrate), Niemann-Pick type C1-like protein (NPC1L1) inhibitors (represented by ezetimibe), bile acid sequestrants (represented by cholestyramine and colesevelam hydrochloride), and niacin compounds also play a crucial role in the prevention, control, and treatment of hyperlipidemia and atherosclerotic heart disease.

[0004] However, the limitations of classic lipid-lowering drugs are gradually emerging. For example, as HMG-CoA reductase inhibitors, statins have some adverse reactions, such as rhabdomyolysis, hepatotoxicity, and cognitive impairment. Clinical trials of niacin compounds have shown that niacin does not significantly improve the risk of atherosclerotic heart disease.

[0005] Currently, new small-molecule lipid-lowering drugs are being researched, including inhibitors of microsomal triglyceride transfer protein (MTP), ATP citrate lyase (ACL) inhibitors, plasma cholesterol ester transfer protein (CETP) inhibitors, and novel peroxisome proliferator-activated receptor agonists. For example, Chinese patent CN110305102A discloses 1,3-benzodioxole natural polyphenolate ester compounds and their pharmaceutical applications. These compounds incorporate a natural polyphenolic acid structure into their structure, using sesamol as a functional fragment. These compounds exhibit significant lipid-lowering activity, simultaneously lowering both triglyceride and cholesterol (TG) levels, while also exhibiting significant anti-lipid peroxidation and antioxidant enhancement. These compounds possess a more potent and broad-spectrum lipid-lowering effect, demonstrating their ability to simultaneously lower TG and TC levels, while also exhibiting anti-lipid peroxidation and antioxidant enhancement.

[0006] Afamelanotide, molecular formula C 78 H 111 N 21 O19 , the structural formula is as follows: .

[0007] Currently, it is used in the adjunctive treatment of erythropoietic protoporphyria (EPP) and vitiligo. Afamelanotide stimulates the melanocortin 1 receptor (MC1R), promoting eumelanin production, alleviating patients' photosensitivity and reducing the risk of UV-induced skin burns and damage. It also regulates immune system function and inhibits autoimmune attacks on melanocytes. Activating the MC1R receptor promotes melanocyte proliferation and melanin synthesis, accelerating pigmentation in vitiligo areas.

[0008] Semaglutide, a GLP-1 receptor agonist, is primarily indicated for the treatment of diabetes and obesity. Semaglutide also indirectly improves blood lipids by delaying gastric emptying, increasing satiety, and reducing calorie intake while directly promoting the decomposition of visceral fat. However, its efficacy in lowering blood lipids alone is limited.

[0009] At present, the research on new and highly effective lipid-lowering drugs is still very necessary, and the research on afamelanotide combined with semaglutide in lipid-lowering is still blank. Summary of the Invention

[0010] The purpose of the present invention is to provide a combination of afamelanotide and semaglutide and its use in the preparation of lipid-lowering drugs, which provides a new direction for the research and development of lipid-lowering drugs and broadens the application field of afamelanotide.

[0011] In order to achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows: In a first aspect, the present invention provides a combination of afamelanotide and semaglutide.

[0012] Afamelanotide, molecular formula C 78 H 111 N 21 O 19 , the structural formula is as follows: .

[0013] Preferably, the weight ratio of the disulfide cyclic peptide compound to semaglutide is 100:0.12-0.2.

[0014] More preferably, it is 100:0.12-0.18, and most preferably, it is 100:0.12-0.164.

[0015] In a second aspect, the present invention provides use of the above combination in the preparation of lipid-lowering drugs.

[0016] Preferably, the lipid-lowering comprises reducing the content of at least one of TC and TG in serum.

[0017] More preferably, the lipid-lowering is to simultaneously reduce the levels of TC and TG in serum.

[0018] In a third aspect, the present invention provides a lipid-lowering drug, the active ingredient of which is the above combination.

[0019] Preferably, the lipid-lowering drug further comprises a pharmaceutically acceptable carrier and / or excipient.

[0020] The phrase "pharmaceutically acceptable carrier" is art-recognized and includes pharmaceutically acceptable materials, components, or vehicles that are suitable for administering the compounds of the present invention to mammals. Carriers include liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials that participate in carrying or transporting the subject substance from one organ or part of the body to another. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation or not deleterious to the subject.

[0021] Some examples of materials that can serve as pharmaceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium methylcellulose, ethylcellulose, and cellulose acetate; tragacanth, malt, gelatin, and talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol, polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol, esters such as ethyl oleate and ethyl laurate, agar, buffers such as magnesium hydroxide and aluminum hydroxide, alginic acid, pyrogen water, isotonic saline, ethanol, phosphate buffer, and other non-toxic, compatible substances used in pharmaceutical formulations. Wetting agents, emulsifiers, and lubricants such as sodium lauryl sulfate and stearates, as well as colorants, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives, and antioxidants can also be present in the composition.

[0022] Examples of pharmaceutically acceptable antioxidants include: water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; oil-soluble antioxidants, such as ascorbyl palmitate, butylated benzoic acid (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and metal complexes, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0023] Suitable pharmaceutically acceptable carriers include, but are not limited to, water, saline solutions (e.g., NaCl), alcohol, gum arabic, vegetable oils, benzyl alcohol, polyethylene glycol, gelatin, sugars (e.g., lactose, amylose, or starch), polyethylene glycol, magnesium stearate, talc, silicic acid, viscous paraffin, aromatic oils, fatty acid esters, methylcellulose, polyvinylpyrrolidone, etc. The pharmaceutical composition can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salt buffers for influencing osmotic pressure, colorants, flavorings, and / or aromatic substances, etc., which do not deleteriously react with the active compound.

[0024] The lipid-lowering drug may also contain a minor amount of a wetting agent, emulsifier, or pH buffer. The lipid-lowering drug may be a liquid solution, suspension, emulsion, tablet, pill, capsule, sustained-release formulation, or powder. The composition may be formulated into a suppository together with conventional binders and carriers (e.g., triglycerides). Oral formulations may include standard carriers such as pharmaceutical grade mannitol, lactose, magnesium stearate, polyvinylpyrrolidone, saccharin sodium, cellulose, magnesium carbonate, and the like.

[0025] According to conventional methods, the lipid-lowering drug can be formulated into a pharmaceutical composition suitable for intravenous administration to humans. When necessary, the composition may further include a stabilizer and a local anesthetic to relieve pain at the injection site.

[0026] Typically, ingredients are supplied separately or mixed together in unit dosage form, for example as dry lyophilized powders or anhydrous concentrates in sealed containers such as ampoules or sachets showing the amount of active agent. When the lipid-lowering drug is to be administered by infusion, it can be dispersed with an infusion bottle containing pharmaceutical grade sterile water, saline or dextrose water. When the lipid-lowering drug is administered by injection, an ampoule of sterile water for injection or saline can be provided so that the ingredients can be mixed before administration.

[0027] The lipid-lowering drugs of the present invention may also include agents that control the release of the compounds of the present invention, thereby providing timed or sustained-release compositions.

[0028] The lipid-lowering drugs of the present invention include formulations suitable for oral, rectal, topical, vaginal and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most appropriate route in any particular case depends on the specific subject, the nature and severity of the condition to which the active ingredient is administered. The pharmaceutical composition can be prepared by any method known in the art of pharmacy.

[0029] The lipid-lowering drugs of the present invention can be administered orally in solid or liquid dosage forms, such as capsules, tablets, lozenges, troches, granules, and powders, or in liquid dosage forms, such as elixirs, syrups, emulsions, dispersions, and suspensions. The active ingredient can also be administered parenterally in sterile liquid dosage forms, such as dispersions, suspensions, or solutions. Other dosage forms that can be used to administer the active ingredient include ointments, creams, drops, transdermal patches, or powders for topical administration; ophthalmic solutions or suspensions, i.e., eye drops, for administration to the eye; sprays or powder compositions for inhalation or intranasal administration; or creams, ointments, sprays, or suppositories for rectal or vaginal administration. Gelatin capsules contain the active ingredient and a powdered carrier, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc. Similar diluents can be used to prepare compressed tablets. Both tablets and capsules can be formulated as sustained-release products to provide sustained release of the drug over several hours. Compressed tablets can be coated with sugar or film to cover any unpleasant taste and protect the tablet from air, or can be enteric-coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration can include coloring agents and flavoring agents to increase patient acceptance. Generally speaking, water, suitable oil, saline, dextrose (glucose) aqueous solution and related sugar solution and glycols such as propylene glycol or polyethylene glycol are the carriers of suitable parenteral solutions. The solution for parenteral administration preferably includes a water-soluble salt of active ingredient, a suitable stabilizer and the buffer substance used as required. Antioxidant such as sodium bisulfite, sodium sulfite or ascorbic acid alone or in combination is a suitable stabilizer. Citric acid and its salt and sodium EDTA can also be used. In addition, parenteral solutions can also include preservatives, such as benzalkonium chloride, methylparaben or propylparaben and chlorobutanol.

[0030] For administration by inhalation, blood lipid lowering drug of the present invention can be easily sent with spray form from pressurized packaging or aerosol.This blood lipid lowering drug can also be sent with the powder form that prepares, and this powder composition can be sucked with the help of blowing into powder inhaler device.Preferred delivery system for sucking is metered dose inhalation (MDI) aerosol, its suspension or the solution in suitable propellant, described propellant is fluorocarbon or hydrocarbon for example.For eye administration, ophthalmic preparation can prepare with the solution or the suspension of the suitable weight percentage of medicine in suitable eye carrier, thereby keep medicine and eye surface contact enough time so that compound penetrates into cornea and the inner area of ​​eyes.

[0031] Useful pharmaceutical dosage forms for administering the lipid-lowering drugs of the present invention include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injections, and oral suspensions.

[0032] As a preferred embodiment of the lipid-lowering drug of the present invention, the dosage form of the lipid-lowering drug is selected from tablets, injections, capsules, granules, suppositories, pills, inhalation preparations, sprays, aerosols, gels, powders, syrups, oral solutions, oral suspensions, oral emulsions, implants, and films.

[0033] When the combination of the present invention is administered stepwise or in combination with other therapeutic agents, the same dosage forms as described above can be used. When the drugs are administered in a physical combination, the dosage form and route of administration should be selected based on the compatibility of the combined drugs. The combination of the present invention can be administered as the sole active ingredient or in combination with a second active ingredient, including active ingredients known to be useful in treating related diseases.

[0034] In the lipid-lowering drug, the mass content of the above combination is 0.01%-10%. In some embodiments, it can be 0.05%-0.1%, 0.1%-1%, 1%-2%, 2%-3%, 3%-4%, 4%-5%, 6%-7%, or 7%-8%.

[0035] In a fourth aspect, the present invention provides a method for lowering blood lipids, comprising administering an effective amount of the aforementioned combination or the aforementioned lipid-lowering drug to an individual. The method can be used in vivo or in vitro. The individual can be a mammal, such as a human.

[0036] The above-mentioned combination or the above-mentioned lipid-lowering drug is a unit dosage form suitable for a single administration of an exact dose. In other embodiments, the amount of the compound is in the range of about 0.001 mg / kg body weight / day to about 1000 mg / kg body weight / day. In other embodiments, the amount of the compound is in the range of about 0.5 mg / kg body weight / day to about 50 mg / kg body weight / day. In some embodiments, the amount of the compound is about 0.001 g / day to about 7 g / day. In other embodiments, the amount of the compound is about 0.002 g / day to about 6 g / day. In other embodiments, the amount of the compound is about 0.005 g / day to about 5 g / day. In other embodiments, the amount of the compound is about 0.01 g / day to about 5 g / day. In other embodiments, the amount of the compound is about 0.02 g / day to about 5 g / day. In other embodiments, the amount of the compound is about 0.05 g / day to about 2.5 g / day. In other embodiments, the amount of the compound is from about 0.1 g / day to about 1 g / day. In other embodiments, dosage levels below the lower limit of the above range may be sufficient. In other embodiments, dosage levels above the upper limit of the above range may be required. In some embodiments, the compound is administered in a single dose once a day. In other embodiments, the compound is administered in multiple doses more than once a day. In some embodiments, the compound is administered twice a day. In other embodiments, the compound is administered three times a day. In other embodiments, the compound is administered four times a day. In other embodiments, the compound is administered more than four times a day. In some embodiments, the individual to whom the pharmaceutical composition is administered is a mammal. In other embodiments, the mammal is a human.

[0037] The beneficial effects of the present invention are: The present invention provides a combination of afamelanotide and semaglutide, and use thereof in the preparation of a lipid-lowering drug. Animal experiments have shown that the combination can significantly reduce the levels of TC and TG in serum, providing a new direction for the research and development of lipid-lowering drugs and broadening the application field of the afamelanotide. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 Figure 2 is the total cholesterol (TC) content of DIO mice in each group.

[0039] Figure 2 Figure 2 shows the triglyceride (TG) content of DIO mice in each group. DETAILED DESCRIPTION The following non-limiting examples are provided to enable those skilled in the art to more fully understand the present invention, but are not intended to limit the present invention in any way. The following is merely an illustrative description of the scope of the present invention, and those skilled in the art may make various changes and modifications to the present invention based on the disclosed content, which should also fall within the scope of protection claimed in this application.

[0040] The present invention will be further described below by way of specific examples. The various chemical reagents used in the examples of the present invention were obtained through conventional commercial channels unless otherwise specified. Unless otherwise specified, the contents described below are all by weight. Unless otherwise specified, it is understood that the experiments were conducted at room temperature.

[0041] Terms and abbreviations: QD: Once a day.

[0042] SC: subcutaneous injection.

[0043] PBS: phosphate-buffered saline.

[0044] Vehicle: control group.

[0045] In the following examples, afamelanotide (also referred to as WP203a in the examples) was obtained from Hangzhou Gutuo Biotechnology Co., Ltd. with a batch number of P231223-A006. Semaglutide was obtained from Zhongpeptide Biochemical Co., Ltd. with a batch number of CX-10-01396.

[0046] Lipid-lowering effect test: 1.1 Experimental Animals Experimental animals: Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. (VRL): C57BL / 6J male mice, 15 weeks old.

[0047] The rearing conditions are as follows: Temperature: 20-26℃; Humidity: 40-70%; Photoperiod: 12 h (7:00 am - 7:00 pm); High-fat diet (HFD): Research Diets, D12492 (Protein: 20% Kcal. Fat: 60%. Kcal. Carbohydrate: 20% Kcal); Drinking water is pure water (used after high-pressure steam sterilization); During the feeding process, the rats were given free access to food and water.

[0048] During the experiment, all animals were housed in IVC cages under the following conditions: temperature: 20-26°C, humidity: 40-70%, photoperiod: 12 h (7:00 am-19:00 pm), drinking water: purified water (autoclaved for use), and feed: high-fat diet (HFD): Research Diets, D12492 (Protein: 20% Kcal, Fat: 60%. Kcal, Carbohydrate: 20% Kcal).

[0049] 1.2 Preparation of dosing solution: Vehicle: PBS (Shanghai Yuanye Biotechnology Co., Ltd., R29076, KR32630A), 0.01 M, pH 6.8.

[0050] Preparation of dosing solution: Weigh an appropriate amount of afamelanotide powder and dissolve it in a solvent to prepare a 40 mg / mL dosing solution. Dissolve the solution using ultrasound. Prepare the test sample every two days and store the dosing solution at 2-8°C.

[0051] 1.3 Animal grouping and drug administration The experimental period lasted 24 days, with drug administration for 21 days. The groups and dosages are shown in Table 2 (experimental week: defined as the week of the first administration of the test article as week 1, with the day being D0). The dosing schedule and dosage are shown in the table below.

[0052]

[0053] Blood collection: On the day of blood collection, animals were fasted for approximately 6 hours. Blood was collected for biochemical analysis. Blood was collected from the submandibular vein. Whole blood was allowed to clot at room temperature for at least 30 minutes, then centrifuged at 1000 g for 10 minutes at 4°C. The supernatant (serum) was transferred to a pre-labeled AXYGEN EP tube and stored at 2-8°C until analysis. TC (total cholesterol) and TG (triglyceride) levels were measured using a Hitachi 7180 (ISE) instrument.

[0054] 1.4 Experimental Results The results are as follows Figure 1-Figure 2 shown.

[0055] Figure 1 The results of total cholesterol testing in animals showed that afamelanotide alone had no significant effect on lowering total cholesterol, while semaglutide alone had a certain effect on lowering total cholesterol. The dosing regimen of Example 1 further enhanced the effect of lowering total cholesterol, while other ratios did not show an enhanced effect.

[0056] Figure 2The results of triglyceride testing in animals showed that neither afamelanotide nor semaglutide alone had a significant effect on lowering triglycerides, while the dosing regimens of Examples 1 and 2 significantly improved the triglyceride-lowering effect, indicating that afamelanotide and semaglutide exhibit a synergistic effect at an appropriate ratio. Among them, the dosing regimen of Example 1 had the best effect.

[0057] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A combination of afamelanotide and semaglutide.

2. The combination according to claim 1, characterized in that The weight ratio of the disulfide cyclic peptide compound to semaglutide is 100:0.12-0.

2.

3. The combination according to claim 2, characterized in that The weight ratio of the disulfide cyclic peptide compound to semaglutide is 100:0.12-0.

18.

4. The combination according to claim 3, characterized in that The weight ratio of the disulfide cyclic peptide compound to semaglutide is 100:0.12-0.

164.

5. Use of the combination according to any one of claims 1 to 5 in the preparation of a lipid-lowering drug.

6. The use according to claim 5, characterized in that The lipid-lowering includes reducing the content of at least one of TC and TG in serum.

7. The use according to claim 6, characterized in that The lipid-lowering method is to simultaneously reduce the levels of TC and TG in serum.

8. A lipid-lowering drug, characterized in that: The active ingredient is the combination according to any one of claims 1 to 5.

9. The drug according to claim 8, characterized in that The lipid-lowering drug further includes a pharmaceutically acceptable carrier and / or excipient.

10. The drug according to claim 9, characterized in that The dosage form of the lipid-lowering drug is selected from tablets, injections, capsules, granules, suppositories, pills, inhalation preparations, sprays, aerosols, gels, powders, syrups, oral solutions, oral suspensions, oral emulsions, implants, and films.

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