阅读说明：本技术 Daphnane型大环二萜类化合物制备及应用 (Preparation and application of Daphnane type macrocyclic diterpenoid compound ) 是由 朱建勇 张宏 潘蓉蓉 张春燕 夏伟 叶颖 李媛 赵亮 张冰冰 罗兰 翟晓翔 于 2020-03-19 设计创作，主要内容包括：本发明涉及药物领域,特别是涉及一种化合物或其药学上可接受的盐、异构体、前药、多晶型物或溶剂化物。本发明提供一种化合物或其药学上可接受的盐、异构体、前药、多晶型物或溶剂化物,所述化合物的化学结构式如式I、或式II所示。本发明所提供的化合物是一种从天然产物中提取的新化合物,所述化合物可以从芫花提取物中提取获得,可以应用在PI3K/Akt/mTOR通路介导的抑制肿瘤细胞(例如,结肠癌细胞)增殖,且表现出非常好的抗增殖促凋亡活性,从而具有良好的产业化前景。(The invention relates to the field of medicines, in particular to a compound or pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof. The invention provides a compound or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, wherein the chemical structural formula of the compound is shown as a formula I or a formula II. The compound provided by the invention is a novel compound extracted from a natural product, can be extracted from a lilac daphne flower bud extract, can be applied to inhibiting the proliferation of tumor cells (such as colon cancer cells) mediated by a PI3K/Akt/mTOR pathway, and shows very good anti-proliferation and apoptosis-promoting activity, thereby having good industrialization prospect.)

1. A compound, or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, having a chemical structural formula as shown in formula I, or formula II:

wherein R is₁、R₂Each independently selected from hydroxy, or R₁、R₂Together with the bridging oxygen atoms form a fused, ternary epoxy group;

R₃is selected from-COR₉Wherein R is₉Is selected from C₁-C₄Alkyl, aryl;

R₄is selected from C_1-10Alkenyl, aryl;

R₅、R₆each independently selected from hydroxy, or R₅、R₆Together with the bridging oxygen atoms form a fused, ternary epoxy group;

R₇is selected from-COR₁₀Wherein R is₁₀Is selected from C₁-C₄Alkyl, aryl;

R₈selected from hydroxy, -COR₁₁Wherein R is₁₁Is selected from C_1-10An alkenyl group.

2. The compound of claim 1, wherein R is₃Is selected from-COR₉Wherein R is₉Is selected from C₁-C₄Alkyl, phenyl;

R₄is selected from C_1-10Dienyl, phenyl;

R₇is selected from-COR₁₀Wherein R is₁₀Is selected from C₁-C₄Alkyl, phenyl;

R₈selected from hydroxy, -COR₁₁Wherein R is₁₁Is selected from C_1-10Dienyl radical, C_1-10A trienyl group.

3. The compound of claim 1, wherein R is₉Selected from butyl, isopropyl, phenyl;

R₄selected from 1, 3-nonadienyl, phenyl;

R₁₀selected from ethyl, phenyl;

R₈selected from the group consisting of hydroxy, 2, 4-decadienoyl, 2,4, 6-decatrienoyl.

4. The compound of claim 1, wherein the isomers are selected from enantiomers, diastereomers, cis-trans isomers, or stereoisomers.

5. The compound of claim 1, wherein the chemical structural formula of the compound is selected from the group consisting of:

6. a process for the preparation of a compound as claimed in any one of claims 1 to 5, which comprises: is prepared from flos Genkwa extract.

7. Use of a compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, in the manufacture of a medicament.

8. The use according to claim 7, wherein the medicament is selected from a PI3K inhibitor, or a PI3K/Akt/mTOR pathway inhibitor;

and/or, the drug is selected from drugs for treating tumors.

9. Use according to claim 8, wherein the tumour is preferably selected from colon cancer, rectal cancer, or melanoma.

10. A pharmaceutical composition comprising a compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof.

Technical Field

The invention relates to the field of medicines, in particular to a compound or pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, wherein the compound can be prepared from lilac daphne flower bud extract.

Background

Colorectal cancer (CRC) is a common gastrointestinal malignancy occurring in the colon region of a human body, and occurs well at the junction between the rectum and the sigmoid colon, with the highest incidence rate in the age group of 40-50 years, and the ratio of male to female is 2-3: 1, it is a malignant tumor disease that seriously threatens human health. Statistical data published in 2014 annual registration of Chinese tumor shows that the morbidity and mortality of colorectal cancer in China are always in an increasing trend, and in the ranking of tumor morbidity, colorectal cancer exceeds gastric cancer for the first time, jumps over the second place of national malignant tumors, and seriously affects the life quality and life health of human beings, so that the research on the aspect related to treatment of colorectal cancer is enhanced, and a new method for treating colorectal cancer is explored, which has important significance.

At present, the effective treatment method for colorectal cancer at home and abroad mainly comprises means of surgical treatment, medicinal chemotherapy, targeted treatment, traditional Chinese medicine treatment and the like. Radical surgery remains the first treatment modality for colon cancer, and the most commonly used drugs for chemotherapy of colorectal cancer are fluorouracil (i.e., 5-fluorouracil and capecitabine), calcium folinate, oxaliplatin and irinotecan. With the molecular pathway and biological process mechanism being continuously understood in the process of tumorigenesis and development, targeted therapeutic drugs are beginning to appear and are gradually applied to clinic. Despite the advent of new technologies and methods for treating colorectal cancer, the therapeutic efficacy remains poor and relapse and metastasis are important causes of patient death. Chemotherapy and targeted drug therapy can appropriately prolong the survival time of cancer patients, but colorectal cancer cannot be effectively cured, and the drugs have adverse reactions and drug resistance in different degrees, so that the development of novel colorectal cancer drugs is accelerated and delayed. The search of key molecular therapeutic targets and the discovery of effective therapeutic drugs based on the targets have important significance for reducing the death rate of colorectal cancer and improving survival prognosis.

Disclosure of Invention

In view of the above-mentioned drawbacks of the prior art, it is an object of the present invention to provide a compound or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, which solves the problems of the prior art.

To achieve the above and other related objects, the present invention provides, in one aspect, a compound having a chemical structural formula as shown in formula I, or formula II, or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof:

wherein R is₁、R₂Each independently selected from hydroxy, or R₁、R₂Together with the bridging oxygen atoms form a fused, ternary epoxy group;

R₃is selected from-COR₉Wherein R is₉Is selected from C₁-C₄Alkyl, aryl;

R₄is selected from C_1-10Alkenyl, aryl;

R₅、R₆each independently selected from hydroxy, or R₅、R₆Together with the bridging oxygen atoms form a fused, ternary epoxy group;

R₇is selected from-COR₁₀Wherein R is₁₀Is selected from C₁-C₄Alkyl, aryl;

R₈selected from hydroxy, -COR₁₁Wherein R is₁₁Is selected from C_1-10An alkenyl group.

In some embodiments of the invention, R₃Is selected from-COR₉Wherein R is₉Is selected from C₁-C₄Alkyl, phenyl;

R₄is selected from C_1-10Dienyl, phenyl;

R₇is selected from-COR₁₀Wherein R is₁₀Is selected from C₁-C₄Alkyl, phenyl;

R₈selected from hydroxy, -COR₁₁Wherein R is₁₁Is selected from C_1-10Dienyl radical, C_1-10A trienyl group.

In some embodiments of the invention, R₉Selected from butyl, isopropyl, phenyl;

R₄selected from 1, 3-nonadienyl, phenyl;

R₁₀selected from ethyl, phenyl;

R₈selected from the group consisting of hydroxy, 2, 4-decadienoyl, 2,4, 6-decatrienoyl.

In some embodiments of the invention, the isomers are selected from enantiomers, diastereomers, cis-trans isomers or stereoisomers.

In some embodiments of the invention, the chemical structural formula of the compound may be selected from:

in another aspect, the present invention provides a process for preparing the above compound, comprising: is prepared from flos Genkwa extract.

In another aspect, the present invention provides the use of a compound as described above, or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, for the manufacture of a medicament.

In some embodiments of the invention, the drug is selected from a PI3K inhibitor, or a PI3K/Akt/mTOR pathway inhibitor;

and/or, the drug is selected from drugs for treating tumors.

In some embodiments of the invention, the tumor is preferably selected from colon cancer, rectal cancer, or melanoma.

In another aspect, the present invention provides a pharmaceutical composition comprising the above compound or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof.

Drawings

FIG. 1 shows a schematic diagram of the isolation scheme of the compounds in the example of the present invention.

FIG. 2 is a graph showing the inhibition curves of DGP-1 on different human colon cancer cells in the present example.

FIG. 3 is a schematic diagram showing the results of the AO/PI staining method for observing that DGP-1 promotes SW620 apoptosis.

FIG. 4 is a diagram illustrating the effect of DGP-1 on SW620 apoptosis detected by flow cytometry in the present invention.

FIG. 5 is a schematic diagram showing the effect of Figure on SW620 apoptosis of DGP-1 detected by Figure flow cytometry in accordance with an embodiment of the present invention, wherein (A) is a cell cycle diagram; (B) the figure is a statistical analysis figure.

FIG. 6 is a graph showing the effect of DGP-1 on the expression of SW 620-related apoptosis proteins in an example of the present invention.

FIG. 7 is a graph showing the effect of DGP-1 on the expression of key proteins and downstream proteins of PI3K/AKT/mTOR pathway in the present invention.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments, and other advantages and effects of the present invention will be apparent to those skilled in the art from the disclosure of the present specification.

The inventor of the invention provides a compound which can be used as a PI3K inhibitor through a large amount of practical researches, can inhibit the proliferation of various colon cancer cell strains, and has obvious inhibition effect on cell PI3K/Akt/mTOR signal transduction, thereby completing the invention.

In a first aspect, the present invention provides a compound, or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, wherein the chemical structural formula of the compound is as shown in formula I or formula II:

wherein R is₁、R₂Each independently selected from hydroxy, or R₁、R₂Together with the bridging oxygen atoms form a fused, ternary epoxy group;

R₃is selected from-COR₉Wherein R is₉Is selected from C₁-C₄Alkyl, aryl;

R₄is selected from C_1-10Alkenyl, aryl;

R₅、R₆each independently selected from hydroxy, or R₅、R₆Together with the bridging oxygen atoms form a fused, ternary epoxy group;

R₇is selected from-COR₁₀Wherein R is₁₀Is selected from C₁-C₄Alkyl, aryl;

R₈selected from hydroxy, -COR₁₁Wherein R is₁₁Is selected from C_1-10An alkenyl group.

Isotopically labeled forms of the compounds of the present invention are also encompassed within the scope of the present invention unless otherwise indicated. For example, in the compounds having the structure of the present invention given above, at least one hydrogen atom is replaced by deuterium or tritium, or at least one carbon is replaced by¹³C-or¹⁴C-enriched carbon, or at least one nitrogen being replaced by¹⁵N-enriched nitrogen substitution.

In the present invention, the term "salt" is to be understood as any form of active compound used by the present invention, wherein said compound may be in ionic form or charged or coupled to a counter ion (cation or anion) or in solution. This definition may also include quaternary ammonium salts and complexes of the active molecule with other molecules and ions, particularly complexes through ionic interactions. This definition includes in particular physiologically acceptable salts, which term is to be understood as being equivalent to "pharmacologically acceptable salts".

In the present invention, the term "pharmaceutically acceptable salt" generally refers to any salt (in general, this means that it is non-toxic, in particular as a result of counterions, non-toxic) that is physiologically tolerable when used in an appropriate manner for therapy, in particular when applied or used in humans and/or mammals. These physiologically acceptable salts may be formed with cations or bases and in the context of the present invention, especially when administered in humans and/or mammals, they are to be understood as being salts formed by at least one compound provided according to the invention, usually an acid (deprotonated), such as an anion, and at least one physiologically tolerated cation, preferably an inorganic cation. In the context of the present invention, salts with alkali metals and alkaline earth metals, and ammonium cations (NH) may be included in particular₄ ⁺) The salts formed may, in particular, include but are not limited toLimited to salts with (mono) or (di) sodium, (mono) or (di) potassium, magnesium or calcium. These physiologically acceptable salts may also be formed with anions or acids, and in the context of the present invention, in particular when administered in humans and/or mammals, they are to be understood as being salts formed by at least one compound provided according to the invention, usually protonated (e.g. on nitrogen), such as a cation and at least one physiologically tolerable anion. In the context of the present invention, salts formed with physiologically tolerable acids, i.e. salts of the particular active compounds with physiologically tolerable organic or inorganic acids, may be included in particular, but not exclusively, with hydrochloric acid, hydrobromic acid, sulfuric acid, methanesulfonic acid, formic acid, acetic acid, oxalic acid, succinic acid, malic acid, tartaric acid, mandelic acid, fumaric acid, lactic acid or citric acid.

The compounds of the present invention represented by formula I above may include enantiomers depending on the presence of chiral centers or isomers (e.g., Z, E) depending on the presence of double bonds. Single isomers, enantiomers, diastereomers or cis-trans isomers and mixtures thereof are within the scope of the invention.

Methods for preparing prodrugs of the named enabling compounds will be known to those skilled in the art, e.g., as disclosed in Krogsgaard-L arsen et al, "Textbook of Drug design and Discovery" (Textbook of Drug design and Discovery) Taylor & Francis, 2002, 4.

In the context of the present invention, the term "solvate" refers generally to any form of substance obtained by non-covalent bonding of an active compound according to the invention to another molecule, usually a polar solvent, and may include in particular, but not exclusively, hydrates and alcoholates, such as methanolate.

In the present invention, the "alkyl group" generally refers to saturated aliphatic groups, which may be straight-chain or branched. E.g. C_1-4Alkyl generally refers to an alkyl group comprising 1, 2, 3, 4 carbon atoms, which specifically may include, but is not limited to, methyl, ethyl, propyl, butyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, and the like.

In the present invention, the "alkenyl group" generally refers to unsaturated aliphatic groups, which may be straight-chain or branched. E.g. C_1-10Alkenyl generally refers to an alkenyl group comprising 1, 2, 3, 4,5,6,7, 8, 9, 10 carbon atoms, which specifically may be, but is not limited to, methyl, ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, and the like. These alkenyl groups may be polybasic, i.e., hydrocarbons containing multiple carbon-carbon double bonds, and may be, for example, nonadienyl, decadienyl, decatrienyl, and the like.

In some preferred embodiments of the invention, R₉Is selected from C₁-C₄Alkyl, phenyl.

In some more preferred embodiments of the invention, R₉Selected from butyl, isopropyl, phenyl.

In some preferred embodiments of the invention, R₄Is selected from C_1-10Dienyl, phenyl.

In some more preferred embodiments of the invention, R₄Selected from 1, 3-nonadienyl and phenyl.

In some preferred embodiments of the invention, R₁₀Is selected from C₁-C₄Alkyl, phenyl.

In some more preferred embodiments of the invention, R₁₀Selected from ethyl and phenyl.

In some preferred embodiments of the invention, R₁₁Is selected from C_1-10Dienyl radical, C_1-10A trienyl group.

In some more preferred embodiments of the invention, 2, 4-decadienoyl, 2,4, 6-decatrienoyl.

In some further preferred embodiments of the invention, the compound may be a compound as shown below:

in a second aspect, the present invention provides a process for the preparation of a compound provided in the first aspect of the present invention, comprising: is prepared from flos Genkwa (Daphne genkwa Sieb. et Zucc) extract.

The preparation method provided by the invention can comprise the following steps: flos Genkwa extract is provided. The flos Genkwa extract is usually flos Genkwa bud extract, and the extract can be alcohol extract. Methods for providing extracts of lilac daphne flower bud should be known to those skilled in the art, for example, in the case of alcoholic extracts, they are generally obtained by extraction with alcoholic solvents, which may typically be ethanol or the like, and the product obtained by extraction may generally be concentrated.

The preparation method provided by the invention can also comprise the following steps: providing extract of flos Genkwa extract. In providing an extract of Daphne genkwa extract, solvents with lower polarity are generally used, which can fully dissolve Daphne type macrocyclic diterpenoids. For example, these solvents may be generally organic solvents, and specifically, petroleum ether, chloroform, methylene chloride, ethyl acetate and the like.

The preparation method provided by the invention can also comprise the following steps: the extract of the genkwa flower extract is separated and purified to provide the compound provided by the first aspect of the invention. Suitable separation and purification methods are known to those skilled in the art, and for example, the lilac daphne flower bud extract can be separated and purified by column chromatography, and specifically, for example, MCI column chromatography, silica gel column chromatography, ODS column chromatography, high performance liquid chromatography, and the like can be used.

In a third aspect, the present invention provides the use of a compound provided in the first aspect of the present invention, or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, in the manufacture of a medicament. As described above, the compound provided by the invention can effectively inhibit the activity or expression level of PI3K, so that the compound has obvious inhibition effect on cell PI3K/Akt/mTOR signaling, can be used as a PI3K inhibitor or a PI3K/Akt/mTOR pathway inhibitor, and can be used for treating various diseases related to PI3K or PI3K/Akt/mTOR pathways. In addition, the compound provided by the invention has a remarkable cytotoxic effect on various tumor cells (such as a colon cancer cell line), can induce G0/G1 cycle arrest of the tumor cells and induce apoptosis of the tumor cells, and can specifically induce apoptosis of the tumor cells through the ratio of bax/bcl-2, activated caspase-3 and caspase-9 and up-regulated PARP, so that the compound can be used as a medicament for treating tumors, specifically colon cancer, rectal cancer, melanoma and the like.

In a fourth aspect, the present invention provides a pharmaceutical composition comprising a compound provided in the first aspect of the present invention or a pharmaceutically acceptable salt, isomer, prodrug, polymorph or solvate thereof, which may further comprise at least one pharmaceutically acceptable carrier.

In the present invention, the composition may include one or more pharmaceutically acceptable carriers, which generally refer to carriers for administration of the therapeutic agent, which do not themselves induce the production of antibodies harmful to the individual receiving the composition, and which are not unduly toxic upon administration. Such carriers are well known to those skilled in the art and are disclosed, for example, in Remington's pharmaceutical Sciences (Mack pub. Co., N.J.1991) in relation to pharmaceutically acceptable carriers. In particular, the carrier may be a combination including, but not limited to, one or more of saline, buffer, glucose, water, glycerol, ethanol, adjuvants, and the like.

In the pharmaceutical composition provided by the invention, the compound can be a single effective component, and can also be combined with other active components to form a combined preparation. The other active component can be other various drugs which can inhibit PI3K activity, inhibit PI3K/Akt/mTOR pathway, or be used for treating tumor. The amount of active ingredient in the composition will generally be a safe and effective amount, which should be adjusted by the person skilled in the art, for example, the amount of the compound and active ingredient of the pharmaceutical composition administered will generally depend on the weight of the patient, the type of application, the condition and severity of the disease, for example, the amount of the compound as an active ingredient administered may be usually 0.1mg to 1mg/kg/day, 0.1mg to 0.2mg/kg/day, 0.2mg to 0.3mg/kg/day, 0.3mg to 0.4mg/kg/day, 0.4mg to 0.5mg/kg/day, 0.5mg to 0.6mg/kg/day, 0.6mg to 0.7mg/kg/day, 0.7mg to 0.8mg/kg/day, 0.8mg to 0.9mg/kg/day, or 0.9mg to 1 mg/kg/day.

The compounds provided herein may be adapted for any form of administration, and may be administered orally or parenterally, for example, by pulmonary, nasal, rectal and/or intravenous injection, and more particularly intradermal, subcutaneous, intramuscular, intraarticular, intraperitoneal, pulmonary, buccal, sublingual, nasal, transdermal, vaginal, oral or parenteral administration. One skilled in the art can select an appropriate formulation according to the mode of administration, for example, a formulation suitable for oral administration may include, but is not limited to, pills, tablets, chewables, capsules, granules, drops or syrups, and the like, and for example, a formulation suitable for parenteral administration may include, but is not limited to, solutions, suspensions, reconstitutable dry preparations or sprays, and for example, a suppository suitable for rectal administration may be the usual.

In a fifth aspect, the invention provides a method of treatment comprising: administering to the subject a therapeutically effective amount of a compound provided by the first aspect of the invention, or a pharmaceutical composition provided by the fourth aspect of the invention.

In the present invention, "subject" generally includes humans, non-human primates, such as mammals, dogs, cats, horses, sheep, pigs, cows, etc., which would benefit from treatment with the formulation, kit or combined formulation.

In the present invention, a "therapeutically effective amount" generally refers to an amount which, after an appropriate period of administration, is capable of achieving the effect of treating the diseases as listed above.

The compound provided by the invention is a novel compound extracted from a natural product, can be extracted from a lilac daphne flower bud extract, can be applied to inhibiting the proliferation of tumor cells (such as colon cancer cells) mediated by a PI3K/Akt/mTOR pathway, and shows very good anti-proliferation and apoptosis-promoting activity, thereby having good industrialization prospect.

The invention of the present application is further illustrated by the following examples, which are not intended to limit the scope of the present application.