WO2011113174A1 - Cytarabine derivatives and uses for anticancer and antitumor thereof - Google Patents

Abstract

Cytarabine derivatives obtained by chemically modifying N4 and O5 position of cytarabine are provided. Methods for synthesizing the cytarabine derivatives, the formulations comprising the cytarabine derivatives and their preparation methods, as well as the uses of the cytarabine derivatives for anticancer and antitumor are also provided.

Description

 Description

Cytarabine derivative and its use in anticancer and antitumor

 The present invention relates to the field of medical technology, in particular to a cytarabine derivative and a synthetic route thereof, and to a cytarabine derivative preparation, a preparation method thereof and a cytarabine derivative and a preparation thereof for anticancer resistance Use in tumors.

Background technique

 Cancer is currently the most important disease that threatens human health. The existing methods of treating cancer include: surgical resection, radiotherapy, chemotherapy or the combination of these methods. Chemotherapy has gained widespread use and has been used in the treatment of many different types of cancer. However, most of the anticancer drugs used in chemotherapy are limited to delaying the deterioration of cancer and prolonging the life of the patient. Although the pathogenesis of various types of cancer varies, they are actually a large group of syndromes with common characteristics. In addition to the strong metabolism and constant differentiation of cancer cells, the physiological difference from normal cells is not large. This is a huge challenge for developing drugs that selectively clear cancer cells without killing normal cells. Another major challenge in the development of anticancer drugs is cancer cell resistance, which is the drug resistance caused by a period of chemotherapy. The used chemotherapy drugs, even if the dose is increased, will no longer play a role in cancer cells. Metastasis of tumor cells often also prevents treatment with chemotherapy. So far, no anticancer drug has been able to cure all cancers. Finding new, highly effective, highly selective, low-toxic, non-resistant, and urgently needed new anticancer drugs remains challenging. Most chemotherapy and anticancer drugs have serious side effects, which can lead to the inability of chemotherapy to continue. Therefore, existing drugs are greatly limited in the treatment of different types of tumors. Therefore, finding new and effective anticancer drugs with high efficiency and low toxicity is still urgently needed to maintain human health.

Cytarabine is an analog of cytidine nucleoside, an inhibitor of DNA polymerase. It can block DNA synthesis, can also be incorporated into DNA, interfere with DNA replication, and can also block the reduction of cytosine nucleotides into Deoxycytosine ¹ 3⁄4 nucleotide (Sylvester, R. Κ·, Fisher, AJ, and Lobell, Μ·, Drug Intelligence & Clinical Pharmacy: Vol. 21, No. 2, pp. 177-180 (1987); Boyer Et al, Novel Cytarabine Monophospate Prodrugs, United States Patent Application Publication, Pub. No.: US 2007/0037774 A 1, (Feb. 15, 2007); Colon-Cesario, Μ·, Wang, J" Ramos, X., Garcia, HG, Davila, JJ, Laguna, J., Rosado, C, and Pena de Ortiz, SJ Neurosci., 26(20): 5524 -5533 (2006)).

 Currently, cytarabine is mainly used for the treatment of acute leukemia. It is the best for acute myeloid leukemia. It is also effective for acute monocytic leukemia and acute lymphoblastic leukemia. It has certain curative effect on malignant lymphoma, lung cancer, digestive tract cancer, head and neck cancer. It has viral keratitis and epidemic. Conjunctivitis and the like also have a certain effect, however, it is ineffective for most solid tumors. The activity of cytarabine is not very high. In order to improve the therapeutic effect, cytarabine is generally combined with other drugs, such as: oxime daunorubicin, all-trans retinoic acid combined with arsenic trioxide, pirarubicin, topography Kang-Etoposide-cyclophosphamide, fludarabine and the like are used in combination. Cytarabine has side effects such as myelosuppression and digestive tract reaction. A few patients may have side effects such as abnormal liver function, fever, and rash (Bolwell, BJ, Cassileth, PA, Gale, RP Leukemia. 2(5): 253-60 ( 1988); Kimby, E., Nygren, P., Glimelius, B. Acta Oncol. 40(2-3): 231-22 (2001); Stamatopoulos, K. Leukemia Research, Volume 22, Issue 8 , pp 759 - 761, (2003); Burnett, AK, Milligan, D., Prentice, AG, Goldstone, AH, McMullin, MF, Hills, RK, Wheatley, K. Cancer. 109(6): 1007-10 (2007)).

Cytarabine is an antimetabolite, which is phosphorylated by deoxycytidine in cells, converted to active cytarabine, and further converted to the corresponding diphosphate and cytarabine. effect. Cytarabine inhibits DNA polymerase by interfering with the deoxycytidine triphosphate required for DNA synthesis, interfering with nucleotide incorporation into DNA, and inhibiting nucleotide reductase, preventing nucleotides from being converted into Deoxynucleotide, but has no significant effect on the synthesis of RNA and protein. It belongs to the cell cycle-specific drug acting on S phase, and is most sensitive to the role of cells in S proliferative phase, and has a G1/S and S/G2 transition period. Also has a role. Immediately after intravenous injection, it disappears from the blood, 40% can pass the blood-brain barrier, and the drug is mainly metabolized in the liver to inactive glucoside, 70% ~ 90% through the kidney. In order to develop new anticancer drugs that are effective against solid tumors such as liver cancer, new drugs for liver targeting must be sought. Obviously, Anti-hepatitis drugs can be an excellent reference. For example, miftine and adefovir (PMEA) have been approved as anti-viral therapeutics for hepatitis B (Starrett, et al., "Synthesis, oral bioavailability determination, and in vitro evaluation of prodrugs of the antiviral agent 9-[2-(phosphonomethoxy)ethyl]adenine (PMEA)," J Med Chem., 37(12): 1857-64 (1994); Shaw, et al, "Pharmacokinextics and Metabolism of Selected Prodrugs of PMEA in Rats, "Drug Metabolism Dis., 25(3): 362-366 (1997); Wacher, VJ, et al" Advanced Drug Delivery Reviews 46:89-102 (2001); Wacher, et al""Active Secretion and Enterocytic Drug Metabolism Barriers to Drug Absorption," Adv. Drug Del. Rev., 46:89-102 (2001); Murono, et al, "Prevention and inhibition of nasopharyngeal carcinoma growth by antiviral phosphonated nucleoside analogs," Cancer Res., 61 (21 ): 7875-7 (2001)). In 2003, scientists of Metabasis Therapeutics, Inc., K. Raja Reddy, Mark D. Erion, Michael C. Matelich, and Joseph J. Kopcho proposed prodrugs of cyclic nucleoside phosphates as anti-hepatocarcinoma treatments (United States Patent 7,214,668; When the compound enters the liver, it is catalyzed by the liver's CYP 3A4 metabolic enzyme to form an acyclic nucleoside derivative and has anticancer activity. In 2007, Metabasis Therapeutics, Inc. filed a new patent application for a cyclic phosphatidyl cyanoside derivative as an anticancer prodrug (Novel Cytarabine Monophospate Prodrugs, United States Patent Application Publication, Pub. No. : US 2007/0037774 A 1, Boyer et al., Feb. 15, 2007; Phosphonic acid based prodrugs of PMEA and its analogues, United States Patent 7,214,668, Reddy, et al. May 8, 2007). The core of these patent applications is the attachment of cyclic phosphate to the -OCH2-ribose position on the ribose ring of cytarabine, the position 05, without modification of the amino group on the cytosine cytosine ring.

 Cytarabine is generally not used to treat liver cancer because its N4 amino group (see Figure 1) is metabolically deactivated and causes toxicity when its nucleoside skeleton structure enters the liver; on the other hand, its glycocalyx structure The 05 hydroxyl group (see Figure 1) must be activated by phosphorylation, and this activation process is too slow in the liver. Summary of the invention

The present invention aims to overcome the deficiencies of the above prior art and provide an efficient, low toxicity, non-resistance, A cytarabine derivative capable of being rapidly activated, and providing a synthetic route of a cytarabine derivative and a preparation method of a cytarabine derivative preparation, and the present invention also provides a cytarabine derivative and a preparation thereof The application of anti-cancer and anti-tumor applications.

 The cytarabine derivative of the present invention, characterized in that the cytarabine derivative is a compound having the following formula (I):

Wherein X is any one of OH, 0-P(0)(OR) ₂ and a phosphate group, and the phosphate group includes a monophosphate group, a diphosphate group, and a triphosphate group; and is 11, Cw ₆ alkyl group, Any one of a cycloalkyl group, a benzyl group, a phenyl group, and an aromatic ring group;

Wherein, A is the second drug-based or functional group in the multi-target drug, and any one of the following formulas is used.

^ is 11, an alkane group, an alkenyl group, an alkynyl group, and an aryl group; R ² is any one of a nitro group, an amino group, a substituted amino group, a halogen atom group, a nitrile group, and an amide group;

Where L is a connected unit, represented by any of the following structural elements:

Wherein n = 0, 1-5; m = 0, 1-18; the curve "one" indicates that the above-mentioned linking unit L is covalently bonded to its adjacent group; the benzene ring or cyclohexane ring in the linking L The two groups above are connected to the ortho, meta or para.

 Each of the groups defined above may be further substituted, and may contain a hetero atom therein.

The cytarabine prodrug derivative of the formula (I) of the present invention includes a representative formula having the following structural formula

为了清晰起见但并非限制本发明， 除另外说明之外， 本发明所使用的所有 科技术语和在本发明领域内技术人员常使用和理解的意义相同。 本发明所引用 的专利申请或已发表的申请及其他论文均属于原始引用并未加修改。 本专利所用"一个"或"一种"或"一类"意指最少一个 /种 /类或一个 /种 /类或一 个 /种 /类以上。

For the sake of clarity, but not limitation of the invention, all technical terms used in the present invention are the same as those which are commonly used and understood by those skilled in the art of the invention. The patent applications or published applications and other papers cited in the present invention are the original references and are not modified. The use of "a" or "an" or "a" or "an"

 As used herein, "alkyl" means various saturated straight-chain, side-chain or cyclic hydrocarbon groups, particularly including small alkyl groups containing ten or ten carbons or less. For example, mercapto, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, isohexyl, heptyl, octyl and decyl groups, etc. Some typical examples in this definition.

 The "alkenyl group" used in the present invention has the same meaning as the above "alkyl group", but it must have at least one carbon-carbon double bond (C=C), so the alkenyl group used in the present invention includes a straight line containing two to ten carbon atoms. A hydrocarbon group having a branched chain or a cyclic group and having at least one carbon-carbon double bond, such as a vinyl group, a propenyl group, a butenyl group, a pentenyl group, and the like.

 The "alkynyl group" used in the present invention is the above alkyl group or gal group and contains at least one carbon-carbon triple bond. Therefore, the alkynyl group includes a linear chain having two to ten carbon atoms and containing at least one carbon-carbon triple bond. A branched or cyclic hydrocarbon or alkynyl group such as ethynyl, propynyl, butynyl and pentynyl.

 "Saturated" in the present invention means that the group does not contain an unsaturated bond, such as a carbon-carbon double bond or a carbon-carbon triple bond; and "unsaturated" means that the group contains one or more carbon-carbon doubles. Key or carbon-carbon triple bond.

 The "cycloalkyl group" used in the present invention is a cyclic hydrocarbon group and preferably a cycloalkyl group having three to eight carbons. Thus cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane and cyclooctane are all typical examples under this definition. The cycloalkyl group contains one or two carbon-carbon double bonds to form a "cycloalkenyl group". The cycloalkyl group may also have an alkyl group, an alkenyl group, an alkynyl group and other groups.

 The "aromatic group" used in the present invention is a cyclic conjugated aromatic system and may contain one or more non-carbon atoms (other than carbon other nitrogen such as nitrogen) such as phenyl, zeoli and pyridine in the ring. Base.

 The "heterocyclic group" which is also commonly used in the present invention means a cyclic group and a compound in which any plurality of atoms are constituted by a covalent bond, and which contains at least one non-carbon atom. Specifically, a heterocyclic group includes a five- and six-membered ring system containing a nitrogen, sulfur or oxygen non-carbon atom such as pyrazole, pyrrole, pyridine or pyrimidine.

The "alkoxy group" in the present invention means an alkyl group-oxidated group formed by linking an oxygen atom to a linear or branched alkyl group. Examples of such alkoxy groups include a decyloxy group, an ethoxy group, a propoxy group or an isopropoxy group and the like. Similarly, "alkylthio" refers to an alkylsulfide group formed by linking a sulfur atom to a linear or branched alkyl group. Examples of such alkylthio groups include sulfonylthio, ethylthio, propylthio or isopropylthio and the like.

 The "halogen atom group" in the present invention is fluorine, chlorine, bromine or iodine.

 The "amino acid" in the present invention means a substituted natural and non-natural amino acid, a pure L- or D-configuration or a racemic mixture, and a group derived from an amino group.

 It is particularly worthy to further clarify that the various substituents defined above also include groups which are further substituted, wherein these new substituents may also contain other groups. For example, a hydrogen atom on an alkyl group or an aromatic group is substituted with an amino group, a halogen or other group to become a new group belonging to each of the above definitions.

The "phosphoric acid" or "phosphate ester" used in the present invention is the highest oxidation state of the pentavalent phosphorus atom to which four oxygen atoms are attached, one oxygen atom is bonded to the phosphorus atom by a double bond, and the two oxygen atoms are bonded to the phosphorus atom by a single bond. The atoms are connected, and the two oxygen atoms may be hydrogen atoms, negative charges or various alkyl groups, aromatic groups, etc. as defined above, such as -P(=0)(0-) ₂ , -P(0) (OR) ₂ . Another oxygen atom on the phosphorus atom is attached to the derivative of the present invention.

 "Multi-targeted drug" in the present invention means that more than one structural unit in a molecule has bioavailability by acting on multiple targets.

 "Multifunctional" in the present invention means that different groups in a molecule have more than one biological function. The "drug-based" or "functional group" in the present invention means that the group is a drug or non-drug group having a biological function. In the present invention, the cytarabine derivative is linked to the aspirin group on the structural formula of cytarabine. Aspirin (also known as acetylsalicylic acid) is a multifunctional drug with antithrombotic, antieclampsia, antidementia, anticataract, anticancer and other features. It is prepared by directly reacting salicylic acid with acetic anhydride. Aspirin is the most widely used antipyretic, analgesic and anti-inflammatory drug in the world and is a standard preparation for comparing and evaluating other drugs. It has an antithrombotic effect in the body, it can inhibit the release of platelets, inhibit the accumulation of platelets, and is clinically used to prevent the onset of cardiovascular and cerebrovascular diseases.

"Linker" in the present invention means that two groups or drug groups are used in the present connection unit. Alternatively, the functional groups may be joined to form a molecule of the invention.

 The "prodrug" in the present invention means a compound in which the cytarabine derivative of the present invention is used in vivo to cleave or increase a biological component formed by a certain structural unit.

 The curve /w" used in the present invention is a commonly used expression in medicinal chemistry, and represents that the group or the drug group or functional group is bonded thereto by a corresponding atom in the formula (I).

 The organic solvents mentioned in the present invention include: benzotriazole hexafluorophosphate-1 P-BO-P, 4-diaminopyridine (DMAP), Ν, Ν-曱Base amide (DMF), dimethyl sulfoxide (DMSO), polyethylene glycol (PEG) and tetrahydrofuran (THF).

 Another technical problem solved by the present invention is to provide a synthetic route of the cytarabine derivative of the general formula (I), the technical solutions of which are as follows:

 1. A synthetic route for cytarabine derivatives, including the following steps:

 (1) Mix salicylic acid with aliphatic diol, then add 3~5 drops of concentrated acid, and heat to 80 °C for 4~12 hours. After the reaction is finished, dry under reduced pressure or pass through the column. The color separation is purified to obtain the first intermediate product (B);

(2) Dissolving the first intermediate product (B) in CH ₂ C1 ₂ or tetrahydrofuran, then adding the acid anhydride compound and DMAP, heating the reflux reaction for 5-12 hours or stirring at room temperature for 24 hours, and obtaining the second intermediate product (C) ) used directly in the next reaction without further purification;

 (3) Dissolving cytarabine, second intermediate product (C), PyBOP and DMAP in DMF, stirring at room temperature for 12-24 hours, and directly purifying the reaction solution by column chromatography to obtain the general formula (I). The cytidine derivative or the reaction solution is poured into water, and extracted with ethyl acetate three times. The ethyl acetate solution is dried over anhydrous sodium sulfate and filtered, and the filtrate is evaporated to dryness, and purified by column chromatography. A cytarabine derivative of the formula (I).

 The aliphatic diol is any of ethylene glycol, butylene glycol, 1,6-hexanediol, 1,8-octanediol, 1,10-nonanediol, and 1,12-dodecanediol. One; the acid anhydride compound is any one of succinic anhydride, phthalic anhydride, glutaric anhydride, and diglycolic anhydride. (Refer to synthetic route 1, 2, 7-24)

2. The synthetic route of cytarabine derivatives, including the following steps: (1) Salicylic acid and sodium carbonate are dissolved in 3-chloropropanol, heated under reflux for 2.5 hours, and the obtained reaction solution is poured into cold water to separate the layers, and the organic layer is washed with NaHC0 ₃ solution to weakly alkaline, and the organic layer is reused. After washing with a small amount of saturated brine, it is dried over anhydrous sodium sulfate, filtered, and the filtrate is evaporated to dryness to give a third intermediate (D) which is directly used for the next reaction;

(2) Dissolving the third intermediate product (D) in CH ₂ C1 ₂ or tetrahydrofuran, then adding the acid anhydride compound and DMAP, stirring the reaction at room temperature for 24 hours, filtering the reaction liquid, and evaporating the filtrate under reduced pressure to obtain a fourth intermediate. The product (E) is used directly in the next reaction;

 (3) Dissolving cytarabine, fourth intermediate (E), PyBOP and DMAP in DMF, stirring at room temperature for 12-24 hours, and directly purifying the reaction liquid by column chromatography to obtain arabinose of general formula (I) The cytidine derivative or the reaction solution is poured into water, and extracted with ethyl acetate three times. The ethyl acetate solution is dried over anhydrous sodium sulfate and filtered, and the filtrate is evaporated to dryness, and purified by column chromatography. A cytarabine derivative of formula (I).

 The acid anhydride compound is any one of succinic anhydride, phthalic anhydride, glutaric anhydride, and diglycolic anhydride. (Refer to synthetic route 3)

 3. The synthetic route of cytarabine derivatives, including the following steps:

 (1) The decyl salicylate and aminopropanol are dissolved in dioxane, stirred at room temperature for 12 hours, the solvent is removed by rotary evaporation, saturated brine is added, and the pH is adjusted to 4-5 with hydrochloric acid, and then extracted with ethyl acetate. Then, the ethyl acetate solution was dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness, and the obtained intermediate product (F) was directly used for the next reaction.

(2) Dissolving the fifth intermediate product (F) in CH ₂ C1 ₂ or tetrahydrofuran, then adding the acid anhydride compound and DMAP, and stirring the reaction at room temperature for 24 hours, filtering the reaction liquid, and evaporating the filtrate under reduced pressure to obtain a crude product. The intermediate product (G) was used directly in the next reaction without further purification;

(3) Dissolving cytarabine, sixth intermediate (G), PyBOP and DMAP in DMF, stirring at room temperature for 12 to 24 hours, and obtaining the reaction solution directly by column chromatography to obtain a product or pour the reaction solution. The mixture was extracted with EtOAc (3 mL), EtOAc (EtOAc)EtOAc. The acid anhydride compound is any one of succinic anhydride, phthalic anhydride, glutaric anhydride, and diglycolic anhydride. (Refer to synthetic route 4, 6)

 4. The synthetic route of cytarabine derivatives, including the following steps:

(1) o-decyloxybenzoic acid and SOCl _{2 are} dissolved in dichloromethane, heated under reflux for 4 hours, and the seventh intermediate product (H) obtained by rotary evaporation to remove solvent and excess SOCl ₂ is used without further purification. Next reaction;

 (2) Dissolving aminopropanol in dichlorosilane, and cooling to 0 °C, adding the seventh intermediate product (H) dissolved in dichloromethane, adding 0.5 hours, and then stirring at room temperature for 5 hours. The solution was acidified to pH = 5, and the solvent was evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated. Solvent, the obtained eighth intermediate product (J) is directly used in the next reaction without purification;

 (3) Dissolving the eighth intermediate product (J) in tetrahydrofuran, then adding the acid anhydride compound and DMAP, stirring the reaction at room temperature for 24 hours, filtering, and evaporating the filtrate under reduced pressure to obtain the ninth intermediate product (K) without purification. Used in the next reaction;

 (4) Dissolving cytarabine, ninth intermediate product (K), PyBOP and DMAP in DMF, stirring at room temperature for 12-24 hours, and directly purifying the product by column chromatography to obtain a product or pouring the reaction solution into water. The mixture was extracted three times with ethyl acetate. The ethyl acetate solution was dried over anhydrous sodium sulfate and filtered, and the filtrate was evaporated to dryness, and purified by column chromatography to give the cytosine derivative of formula (I).

The acid anhydride compound is any one of succinic anhydride, phthalic anhydride, glutaric anhydride, and diglycolic anhydride. (Refer to Synthetic Route ⁵ )

 5. The synthetic route of cytarabine derivatives, including the following steps:

(1) Adding an aliphatic dibasic acid to S0C1 ₂ , adding DMF, heating under reflux, and reacting for 3 hours, and spinning off S0C1 ₂ , the obtained intermediate product (M) is directly used for the next reaction without further purification;

 (2) Dissolving o-nitroaniline in benzene and adding pyridine, adding dropwise to the tenth intermediate dissolved in benzene

In (M), the reaction is carried out for 4 h, the solvent is distilled off, the reactant is dissolved in water, and the pH is adjusted to be acidic. Filtration, washing with water, dissolving the precipitate in isopropanol, recrystallizing, and separating the eleventh intermediate (N) of the solid;

(3) The cytarabine, the eleventh intermediate product (N), PyBOP and DMAP of the solid were dissolved in DMF, stirred at room temperature for 12 hours, and the reaction liquid was poured into water to precipitate a solid, which was purified by column chromatography. A cytarabine derivative of the formula (I) is obtained.

The aliphatic dibasic acid is any one of oxalic acid, succinic acid, 1,6-hexanediol, and 1,8-octanedioic acid. (Refer to Synthetic Route ²⁵ )

 6. The synthetic route of cytarabine derivatives, including the following steps:

(1) o-Nitrobenzoic acid and SOC2 _{2 were} dissolved in DMF, and refluxed for 2 hours, and the twelfth intermediate product (S) obtained was used in the next step without further purification.

 (2) Dissolving the twelfth intermediate product (S) in tetrahydrofuran, adding dropwise to the aqueous solution of the amino acid compound, and simultaneously adding an aqueous sodium hydroxide solution to control the pH = 8 to 9, and controlling the temperature at about 10 ° C for 30 minutes. After the completion of the dropwise addition, the mixture was stirred for 40 minutes, and the pH was adjusted to 3, and the THF was evaporated to dryness. a thirteenth intermediate product (T); the amino acid compound is alanine or aminocaproic acid; (3) dissolving cytarabine, thirteenth intermediate (T), PyBOP and DMAP in DMF, stirring at room temperature After 12 hours, the reaction mixture was poured into water, and the mixture was combined with EtOAc EtOAc EtOAc. ) ;

 (4) The fourteenth intermediate product (U) is further dissolved in tetrahydrofuran, catalyzed by hydrogenolysis, stirred, filtered, and the filtrate is spin-dried to obtain a highly pure cytarabine derivative of the formula (I). (Refer to the synthetic route 26,

27)

 7. The synthetic route of cytarabine derivatives, including the following steps:

 (1) Dissolving nitrobenzoic acid or nicotinic acid, hexanoicol, PyBOP and DMAP in DMF, stirring at room temperature for 12 hours, pouring the reaction solution into water, extracting three times with ethyl acetate, and using anhydrous sodium sulfate for ethyl acetate. Drying, filtering, and evaporating the filtrate under reduced pressure, and purifying by column chromatography to obtain the fifteenth intermediate product (V);

(2) dissolving the fifteenth intermediate product (V) in tetrahydrofuran, and then adding phthalic anhydride and DMAP, heating under reflux for 12 hours, the obtained sixteenth intermediate product (W) was directly used for the next reaction without further purification;

 (3) Dissolving cytarabine, the sixteenth intermediate product (W), PyBOP and DMAP in DMF, stirring at room temperature for 12 hours, pouring the reaction solution into water, extracting three times with ethyl acetate, and using anhydrous sulfuric acid for ethyl acetate solution. The sodium is dried, filtered, and the filtrate is evaporated to dryness under reduced pressure, and then purified by column chromatography to obtain the 17th intermediate product (Y);

 (4) The seventeenth intermediate product (Y) is further dissolved in tetrahydrofuran, catalyzed by hydrogenolysis, stirred, filtered, and the filtrate is spin-dried to obtain a highly pure cytarabine derivative of the formula (I). (Refer to synthetic route 28, 29)

 A method for preparing a cytarabine derivative preparation of the present invention, which is characterized in that:

 (1) dissolving the cytarabine derivative of the formula (I) into water, physiological saline, aqueous cyclodextrin solution, water-soluble organic solvent, nonionic surfactant, water-soluble lipid, fatty acid, Preparing a preparation solution by combining a solvent of any one or more of a fatty acid ester and a phospholipid;

 (2) The preparation solution is further diluted with physiological saline or glucose injection to prepare a cytarabine derivative.

 The organic solvent may be any one of ethanol, propylene glycol, glycerin, glyceride, polyethylene glycol (PEG), hydrazine, fluorenyl-dimercaptocarboxamide (DMF) and dimercaptosulfoxide (DMSO). Or a combination of solvents.

 The cytarabine derivative preparation of the present invention is characterized in that it is a product prepared by the above-described preparation method of a cytarabine derivative preparation.

 Use of the cytarabine derivative of the invention for anticancer and antitumor. The cytarabine derivative of formula (I) can be used to treat or alleviate cancer in a certain tissue or organ. Cancer includes, but is not limited to, leukemia, solid tumors, lung cancer, colon cancer, liver cancer, central nervous system tumors, ovarian cancer, and kidney cancer.

Use of the cytarabine derivative preparation of the present invention for anticancer and antitumor. Cancers include leukemia, solid tumors, lung cancer, colon cancer, liver cancer, central nervous system tumors, ovarian cancer, and kidney cancer. The cytarabine derivative preparation of the invention can also be used in combination with other chemotherapeutic drugs in anti-tumor chemotherapy, and can be combined with the present invention The anti-tumor drugs used in combination with the preparation of cytarabine derivatives include, but are not limited to, alkylating agents, plant alkaloids, antibacterial antitumor sulfonamides, platinum drugs, anti-metabolisms and other known antibiotics. Cancer drugs. The combination therapy referred to in the present invention includes the use of at least one cytarabine derivative exemplified in the present invention.

 The pharmaceutical preparation is prepared by using the cytarabine derivative of the present invention as a component, and can be administered orally or parenterally. The parenteral administration referred to herein means subcutaneous, intravascular, intraarterial, intrauterine, intraatrial, intrasynovial, intrasternal injection or instillation.

 The present invention contemplates the design of cytarabine derivatives using new design techniques. The reason why cytarabine (refer to Figure 1) cannot be used to treat liver cancer is that on the one hand, when its nucleoside skeleton structure enters the liver, its N4 amino group is metabolically deactivated and causes toxicity, and on the other hand, its glycocalyx structure The 05 hydroxyl group must be activated by phosphorylation, and this activation process is too slow in the liver.

 The cytarabine derivative of the present invention is designed by chemically modifying the position of N4, 05 to prevent the N4 amino group from being metabolically deactivated and causing toxicity, allowing the 05 hydroxyl group to be easily activated by phosphorylation, and the aspirin multifunctional drug is introduced into A. In the prodrug molecule of cytarabine, a novel anti-tumor prodrug derivative was designed to increase its anti-solid tumor activity by increasing its drug function. The cytarabine derivatives of the present invention have multi-target versatility, and the main beneficial effects are: increased bioavailability, reduced multi-drug resistance (multi-target design technique), increased solubility, and increased ester solubility. The present invention provides various synthetic routes of the cytarabine derivative of the general formula (I), a cytarabine derivative preparation and a preparation method thereof, and proves the cytarabine of the present invention by a large amount of experimental data. The use of derivatives in anti-cancer and anti-tumor applications.

DRAWINGS

 Figure 1 is a schematic diagram showing the structure of cytarabine and the position of modification of N4, 05.

 Fig. 2 is a view showing the structure of a representative cytarabine derivative of the present invention.

Fig. 3 is a graph showing the drug concentration-inhibition rate of the cytosine derivative of the present invention inhibiting the BEL-7402 liver cancer cell line. detailed description The synthetic routes of some representative cytarabine derivatives of the present invention are listed below, and other cytarabine derivatives in the present invention are synthesized by the same or similar methods.

 Synthesis of the first intermediate product B1 (Scheme 1): Dissolve salicylic acid (13. 8 g, 100 mmol) in 40 ml of butanediol, then add 5 drops of concentrated sulfuric acid, reflux for 4 hours, and reduce after the reaction. After evaporation to dryness, the obtained first intermediate product B1 is directly used for the next reaction.

 Synthesis of second intermediate C1 (Scheme 1): The first intermediate B1 (2.1 g, 100 mmol) was dissolved in 10 mL of tetrahydrofuran, then succinic anhydride (1.0 g, 100 mmol) and DMAP (1.2 g, 100 mmol), the reaction was stirred at room temperature for 24 hours, and the reaction mixture was filtered, and the filtrate was evaporated to dryness.

Synthesis of cytosolic derivative 1 (Scheme 1): cytarabine (2.43 g, 100 mmol), second intermediate CI (3.1 g, 100 mmol), PyBOP (5.7 g, 110 mmol) and DMAP (0.12 g, 10 mmol) was dissolved in DMF (10 ml) and stirred at room temperature for 24 hours. The reaction solution was purified by column chromatography (silica gel, solvent: dichloromethane / decyl alcohol = 10/1) to give cytidine derivative 1 (42 mg). LC (UV 254 nm) purity 95%. LC-MS m/z 536 [M + H]+ (Molecular Formula 0 ₂₄ Η ₂₉ Ν ₃ Ο _π , Molecular Weight 535 ) ; ^ΝΜΚίόΟΟ MHz, DMSO-d ₆ ) δ 10.90 (s, 1H) , 10.56 (s, 1H), 8.04 (d, 1H), 7.79 (d, 1H), 7.54 (t, 1H), 7.15 (d, 1H), 6.99 (m, 2H), 6.05 (d, 1H), 5.48 (t, 2H), 5.06 (t, 1H), 4.34 (t, 2H), 4.09 (t, 3H), 3.62 (t, 1H), 3.33 (t, 1H), 2.69 (d, 2H), 2.58 ( d, 2H), 2.50 (s, 2H), 1.78 (d, 2H), 1.73 (d, 2H).

 Synthesis of the first intermediate product B2 (Scheme 2): Dissolve salicylic acid (13. 8 g, 100 mmol) in 40 ml of butanediol, then add 5 drops of concentrated sulfuric acid, reflux for 4 hours, and reduce after the reaction. It was evaporated to dryness, and the obtained first intermediate B2 was directly used for the next reaction.

 Synthesis of second intermediate C2 (Scheme 2): The first intermediate B2 (2.1 g, 100 mmol) was dissolved in 10 mL of tetrahydrofuran, then glutaric anhydride (1.14 g, 100 mmol) and DMAP (1.2 g) , 100 mmol), the reaction was stirred at room temperature for 24 hours. The reaction mixture was filtered, and the filtrate was evaporated to dryness under reduced pressure, and the obtained intermediate product C2 was directly used for the next reaction.

Synthesis of cytarabine derivative 2 (Scheme 2): cytarabine (2.43 g, 100 mmol), second intermediate C2 (3.24 g, 100 mmol), PyBOP (5.7 g, 110 mmol) and DMAP (0.12 g, 10 mmol) was dissolved in DMF (10 mL). Stir at room temperature for 24 hours. The reaction solution was purified by column chromatography (silica gel, solvent: dichloromethane / decyl alcohol = 10/1) to give cytidine derivative 2 (93.9 mg), LC (UV 254 nm) purity >95 %. LC-MS m/z 550 [M + H]+ (molecule Formula C ₂₅ H ₃₁ N ₃ O _u , molecular weight 549) synthetic route

物 D3的合成 (路线 3 (31.8 g, 300 mmol )溶于 3-氯丙醇（180 ml) 中， 加热回流 2.5小时。 反应液倒入冷 水中分层， 用 NaHC0₃溶液洗涤有机层至弱碱性， 少量饱和盐水洗 涤后， 加入无水硫酸钠干燥， 过滤， 滤液减压蒸干， 第三中间产物 D3 直接用 于下一步反应。 Third intermediate production

Synthesis of material D3 (Route 3 (31.8 g, 300 mmol) dissolved in 3-chloropropanol (180 ml), heated under reflux for 2.5 hours. The reaction solution was poured into cold water to separate layers, and the organic layer was washed with NaHC0 ₃ solution to weak After washing with a small amount of saturated brine, dried over anhydrous sodium sulfate and filtered, and the filtrate evaporated to dryness.

Synthesis of the fourth intermediate E3 (Scheme 3): The third intermediate D3 (1.96 g, 100 mmol) was dissolved in 10 mL of tetrahydrofuran, then succinic anhydride (1.0 g, 100 mmol) and DMAP (1.2 g, 100 mmol), the reaction was stirred at room temperature for 24 hours, the reaction solution was filtered, and the filtrate was evaporated to dryness.

Synthesis of cytosolic derivative 3 (Scheme 3): cytarabine (2.43 g, 100 mmol), fourth intermediate E3 ( 2.96 g, 100 mmol), PyBOP (5.7 g, 110 mmol) and DMAP (0.12 g, 10 mmol) was dissolved in DMF (10 ml) and stirred at room temperature for 24 h. The reaction solution was purified by column chromatography (silica gel, solvent: dichloromethane / decyl alcohol = 10/1) to give cytidine derivative 3 (20 mg). LC (UV 254 nm) purity 94%. LC-MS m/z 522 [M + H] + (M. C ₂₃ H ₂₇ N ₃ 0 s, molecular weight 521). Combined

Synthesis of the fifth intermediate product F4 (Scheme 4): decyl salicylate (20 g, 130 mmol), aminopropanol (15 g, 200 mmol) dissolved in dioxane (50 ml), stirred at room temperature 12 hour. The solvent was removed by rotary evaporation, saturated brine was added, and pH was adjusted to 4-5 with hydrochloric acid, and extracted with ethyl acetate ten times. The ethyl acetate solution was dried over anhydrous sodium sulfate and filtered, and the filtrate was evaporated to dryness.

Synthesis of the sixth intermediate G (Scheme 4): The fifth intermediate F4 (1.95 g, 100 mmol) was dissolved in 10 mL of THF, then succinic anhydride (1.3 g, 130 mmol) and DMAP (1.56 g, The reaction mixture was stirred at room temperature for 24 hours, and the reaction mixture was filtered, and the filtrate was evaporated to dryness.

Synthesis of cytarabine derivative 4 (Scheme 4): cytarabine (2.43 g, 100 mmol), sixth intermediate G (2.95 g, 100 mmol), PyBOP (5.7 g, 110 mmol) and DMAP (0.12 g, 10 mmol) was dissolved in DMF (10 ml) and stirred at room temperature for 24 h. The reaction solution was purified by column chromatography (silica gel, developing solvent: methylene chloride / decyl alcohol = 10/1) to obtain cytarabine derivative 4 (75 mg), LC (UV 254 nm) purity 92% . LC-MS m/z 521 [M + H] + (M. C ₂₃ H ₂₈ N ₄ O ₁₀ , molecular weight 520). Synthetic route 5:

The synthesis route of the seventh intermediate product H5 (Scheme 5): o-decyloxybenzoic acid (20 g, 131.6 mmol), S0C1 ₂ (15 ml), dissolved in dichloromethane (50 ml), and heated to reflux for 4 hours. The solvent and excess SOCl ₂ were removed by rotary evaporation to give the next intermediate product Η5 which was used in the next step without further purification.

Synthesis route of the eighth intermediate product J5 (Route 5): Aminopropanol (12. 4 g, 164 mmol) was dissolved in dichloromethane (70 ml), and dissolved in dichlorohydrazine when cooled to 0 °C. The seventh intermediate product H5 (14 g, 82 mmol) of alkane (10 ml) was added dropwise over 0.5 hour. Then, the mixture was stirred at room temperature for 5 hours. Concentrated hydrochloric acid was acidified to ρ Η = 5 and the solvent was removed by rotary evaporation. The product was dissolved in a small amount of water and extracted with EtOAc. EtOAc was evaporated. .

Synthesis route of ninth intermediate product Κ5 (Scheme 5): The eighth intermediate product J5 (2.09 g, 10 mmol) was dissolved in 10 ml of THF, then succinic anhydride (1.00 g, 10 mmol) and DMAP (1.2 g) were added. The mixture was stirred at room temperature for 24 hours, filtered, and the filtrate was evaporated to dryness, and the obtained ninth intermediate product Κ5 was used for the next reaction without purification.

Synthesis of cytarabine derivative 5 (Scheme 5): cytarabine (2.43 g, 10 mmol), ninth intermediate Κ5 (3.99 g, 10 mmol), PyBOP (5.7 g, 11 mmol) and DMAP ( 0.12 g, 1 mmol) was dissolved in DMF (10 ml) and stirred at room temperature for 24 h. The reaction solution was purified by column chromatography (silica gel, solvent: dichloromethane / decyl alcohol = 10/1) to obtain cytarabine derivative 5 (38.7 mg); LC (UV 254 nm) purity >90% . LC-MS m/z 535 [M + H] + (M. C ₂ 4H _{3 .} N ₄ O ₁₀ , molecular weight 534). Synthetic route 6:

Synthesis of the fifth intermediate F6 (Scheme 6): decyl salicylate (20 g, 130 mmol), aminopropanol (15 g, 200 mmol) dissolved in dioxane (50 ml), stirred at room temperature 12 hour. The solvent was removed by rotary evaporation, saturated brine was added, and pH was adjusted to 4-5 with hydrochloric acid, and extracted with ethyl acetate ten times. The ethyl acetate solution was dried over anhydrous sodium sulfate and filtered, and the filtrate was evaporated to dryness.

Synthesis of the sixth intermediate G6 (Scheme 6): The fifth intermediate F6 (1.95 g, 10 mmol) was dissolved in 10 mL of THF then glutaric anhydride (1. 14 g, 10 mmol) and DMAP (1.2 g, 10) Methyl acetate was reacted at room temperature for 24 hours, filtered, and the filtrate was evaporated to dryness.

Synthesis of cytarabine derivative 6 (Scheme 6): cytarabine (2.43 g, 10 mmol), sixth intermediate G6 (3.99 g, 10 mmol), PyBOP (5.7 g, 11 mmol) and DMAP (0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction solution is purified by column chromatography (silica gel, developing solvent: dichlorodecane / decyl alcohol = 10/1), cytarabine derivative 6 (82.2 mg), LC (UV 254 nm) purity >95%). LC-MS m/z 535 [M + H] + (M. C ₂ 4H _{3 .} N ₄ O ₁₀ , molecular weight 534). synthetic route

Synthesis of the first intermediate product B7 (Scheme 7): Salicylic acid (27.6 g, 200 mmol) was dissolved in 70 g of ethylene glycol, then 3 ml of concentrated sulfuric acid was added dropwise, and the reaction was refluxed for 4 hours. It was evaporated to dryness under reduced pressure, and the obtained intermediate product B7 (ethylene glycol salic acid) was directly used for the next reaction.

Synthesis of second intermediate C7 (Scheme 7): first intermediate B7 (ethylene glycol salicylate) (2.7 g, 15 mmol), glutaric anhydride (1.7 g, 15 mmol) and DMAP (0.2 g, 1.5 mmol) was dissolved in CH ₂ C1 ₂ (30 ml) and heated to reflux for 5 hours. The obtained second intermediate C7 was used in the next step without further purification.

Synthesis of cyanosine derivative 7 (Scheme 7): cytarabine (2.5 g, 10 mmol), second intermediate C7 (3 g, 10 mmol), PyBOP (5.2 g, 10 mmol) and DMAP ( 0.2 g, 2 mmol) dissolved in DMF (25 mL). The reaction solution was suspended with water, extracted with ethyl acetate, and the extract was purified by column chromatography (silica gel, solvent: dichloromethane / decyl alcohol = 10/1) to obtain cytarabine derivative 7 (23.7 Mg ) , LC (UV 254 nm ) purity 95%. LC-MS m/z 522 [M + H] + (Molecular formula C^H^NsOn, molecular weight 521). synthesis

Synthesis of the first intermediate product B8 (Scheme 8): Salicylic acid (27.6 g, 200 mmol) was dissolved in 70 g of ethylene glycol, then 3 ml of concentrated sulfuric acid was added dropwise, and the reaction was refluxed for 4 hours. It was evaporated to dryness under reduced pressure, and the obtained first intermediate product B8 (ethylene glycol salic acid) was directly used for the next reaction.

Synthesis of the second intermediate C8 (Scheme 8): first intermediate B8 (ethylene glycol salicylate) (8.7 g, 47 mmol), succinic anhydride (4.8 g, 48 mmol) and DMAP (0.6 g, 5 mmol) was dissolved in CH ₂ C1 ₂ (50 ml) and heated to reflux for 5 hours. The obtained second intermediate C8 was used in the next step without further purification.

Synthesis of cytosolic derivative 8 (Scheme 8): cytarabine (2.5 g, 10 mmol), second intermediate C8 (5 g, 15 mmol), PyBOP (5.2 g, 10 mmol) and DMAP ( 0.2 g, 2 mmol) was dissolved in DMF (25 ml) and stirred at room temperature for 12 h. The reaction solution was suspended with water, extracted with ethyl acetate, and the extract was purified by column chromatography (silica gel, solvent: dichloromethane / decyl alcohol = 10/1) to obtain cytarabine derivative 8 (298.3 Mg ) , LC (UV 254 nm ) purity 95%. LC-MS m/z 508 [M + H] ⁺ (Molecular formula C ₂₂ H ₂₅ N ₃ 0 „, molecular weight 507 ).

Synthesis of the first intermediate product B9 (Scheme 9): Mix salicylic acid (13. 8 g, 100 mmol) with 1,6-hexanediol (23.6 g, 200 mmol), then add 5 drops of concentrated sulfuric acid and heat to After reacting at 80 ° C for 5 hours, after completion of the reaction, purification was carried out by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) to obtain the first intermediate product B9.

Synthesis of the second intermediate C9 (Scheme 9): The first intermediate B9 (2.4 g, 10 mmol) was dissolved in THF (30 ml), then glutaric anhydride (1.6 g, 13 mmol) and DMAP (1.7 g, 13 mmol), heated to reflux for 12 hours. The obtained second intermediate C9 was used in the next step without further purification.

Synthesis of cytarabine derivative 9 (Scheme 9): cytarabine (2.4 g, 10 mmol), second intermediate C9 (3.5 g, 10 mmol), PyBOP (5.7 g, 11 mmol), and DMAP ( 0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water and extracted with EtOAc EtOAc EtOAc EtOAc EtOAc. Sterol = 10/1) gave cytarabine derivative 9 (67.8 mg). The purity of LC (UV 254 nm) was 95%. LC-MS m/z 578 [M + H] ⁺ (Molecular formula C ₂₇ H ₃₅ N ₃ O _u , molecular weight 577). Synthetic route 10:

Synthesis of first intermediate product BIO (Scheme 10): Mix salicylic acid (6.9 g, 50 mmol) with 1,8-octanediol (14. 6 g, 100 mmol), then add 5 drops of concentrated sulfuric acid and heat to After reacting at 80 ° C for 12 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) afforded the first intermediate product B10.

Synthesis of the second intermediate C10 (Scheme 10): The first intermediate B10 (1.33 g, 5 mmol) was dissolved in THF (100 ml), then glutaric anhydride (0.74 g, 6.5 mmol) and DMAP (0.79 g) were added. , 6.5 mmol), heated to reflux for 12 hours. The obtained second intermediate C10 was used in the next step without further purification.

Synthesis of cytarabine derivative 10 (Scheme 10): cytarabine (1.2 g, 5 mmol), second intermediate C10 (1.9 g, 5 mmol), PyBOP (3.1 g, 5 mmol), and DMAP (60 mg, 0.5 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 15/1) to give cytarabine derivative 10 (44.7 mg). LC (UV 254 nm) purity 96%. LC-MS m/z 564 [M + H] + (Molecular formula C29H39N30H, molecular weight 605). Synthetic route 11:

第一中间产物 Bll的合成（路线 11)： 将水杨酸（13.8g, 100 mmol)与 1,10- 癸二醇（34.8 g, 200 mmol) 混合， 然后滴加 5滴浓硫酸， 加热至 80°C反应 12 个小时， 反应结束后， 通过柱层析色普提纯（硅胶， 展开剂： 石油醚 /乙酸乙酯 =5/1) , 得到第一中间产物 Bll。

Synthesis of first intermediate product B11 (Scheme 11): Mix salicylic acid (13.8 g, 100 mmol) with 1,10-decanediol (34.8 g, 200 mmol), then add 5 drops of concentrated sulfuric acid and heat to After reacting at 80 ° C for 12 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) gave the first intermediate product B11.

Synthesis of the second intermediate C11 (Scheme 11): The first intermediate Bll (2.9 g, 10 mmol) was dissolved in THF (100 ml), then glutaric anhydride (1.4 g, 12 mmol) and DMAP (1.5 g) , 12 mmol), heated to reflux for 12 hours. The second intermediate CI 1 obtained was used directly in the next reaction without further purification.

Synthesis of cytarabine derivative 11 (Scheme 11): cytarabine (2.4 g, 10 mmol), second intermediate C11 (4.1 g, 10 mmol), PyBOP (5.7 g, 11 mmol) and DMAP (0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction solution was poured into water, and the mixture was extracted with EtOAc EtOAc EtOAc EtOAc EtOAc. /sterol = 25/1), cytarabine derivative 11 (88.8 mg) was obtained. LC (UV 254 nm) purity 96%. LC-MS m/z 634 [M + H] + (Molecular formula C ₃₁ H4 ₃ N ₃ O _u , molecular weight 633 ).

 Synthesis of the first intermediate product B12 (Scheme 12): Mix salicylic acid (3.45 g, 25 mmol) with 1,12-dodecanediol (10.1 g, 50 mmol), then add 5 drops of concentrated sulfuric acid, and heat After reacting for 5 hours at 80 ° C, after completion of the reaction, purification was carried out by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) to obtain a first intermediate product B12.

 Synthesis of the second intermediate C12 (Scheme 12): The first intermediate B12 (0.966 g, 3 mmol) was dissolved in THF (30 ml), then glutaric anhydride (0.455 g, 3.9 mmol) and DMAP (0.475 g) were added. , 3.9 mmol), heated to reflux for 12 hours. The obtained second intermediate C12 was used in the next step without further purification.

, 分子量 661 ) 。 合成路线 13： Synthesis of cytosolic derivative 12 (Scheme 12): cytarabine (0.729 g, 3 mmol), second intermediate C12 ( 1.308 g, 3 mmol), PyBOP ( 1.848 g, 3.3 mmol ) and DMAP (0.06 g, 0.5 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 15/1) to give cytarabine derivative 12 (20.9 mg). LC (UV 254 nm) purity 80%. LC-MS m/z 662 [M + H]+ (Molecular Formula

, molecular weight 661). Synthetic route 13:

Synthesis of first intermediate product B13 (Scheme 13): Mix salicylic acid (3.45 g, 25 mmol) with 1,6-hexanediol (5.9 g, 50 mmol), then add 5 drops of concentrated sulfuric acid and heat to After reacting at 80 ° C for 5 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) afforded the first intermediate product B13.

Synthesis of the second intermediate C13 (Scheme 13): The first intermediate B13 (0.65 g, 2.74 mmol) was dissolved in THF (30 ml), then diglycol (0.38 g, 3.29 mmol) and DMAP (0.4 g, 3.29 mmol), heated to reflux for 12 hours. The obtained second intermediate C13 was used in the next reaction without further purification.

Synthesis of cytosolic derivative 13 (Scheme 13): cytarabine (0.67 g, 2.74 mmol), second intermediate C13 (0.97 g, 2.74 mmol), PyBOP (1.5 g, 2.9 mmol) and DMAP (0.05 g, 0.41 mmol) was dissolved in DMF (10 ml) and stirred at room temperature for 12 h. The reaction mixture was poured into water and extracted with EtOAc EtOAc EtOAc EtOAc. Sterol = 15/1) to give cytarabine derivative 13 (50.5 mg). LC (UV 254 nm) purity 91%. LC-MS m/z 580 [M + H] + (Molecular formula C ₂₆ H ₃₃ N ₃ 0 ₁₂ , molecular weight 579 ). Synthetic Road 14:

Synthesis of first intermediate product B14 (Scheme 14): Mix salicylic acid (6.9 g, 50 mmol) with 1,8-octanediol (14. 6 g, 100 mmol), then add 5 drops of concentrated sulfuric acid and heat to After reacting at 80 ° C for 12 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) afforded the first intermediate product B14.

Synthesis of second intermediate C14 (Scheme 14): The first intermediate B14 (0.48 g, 1.8 mmol) was dissolved in THF (30 ml) then diglycol (0.25 g, 2.2 mmol) and DMAP (0.27) g, 2.2 mmol), heated to reflux for 12 hours. The obtained second intermediate C14 was used in the next step without further purification.

Synthesis of cytosolic derivative 14 (Scheme 14): cytarabine (0.44 g, 1.8 mmol), second intermediate C14 (0.69 g, 1.8 mmol), PyBOP (0.94 g, 1.8 mmol) and DMAP (0.022 g, 0.18 mmol) was dissolved in DMF (5 mL). The reaction mixture was poured into water and extracted with EtOAc EtOAc EtOAc EtOAc. Sterol = 15/1) to give cytarabine derivative 14 (22.9 mg). LC (UV 254 nm) purity 85%. LC-MS m/z 608 [M + H] + (M. C ₂₈ H ₃₇ N ₃ 0 ₁₂ , molecular weight 607).

 Synthesis of first intermediate product B15 (Scheme 15): Mix salicylic acid (13.8 g, 100 mmol) with 1,10-decanediol (34.8 g, 200 mmol), then add 5 drops of concentrated sulfuric acid, and heat to 80 ° C was reacted for 12 hours, and after completion of the reaction, purification was carried out by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) to obtain the first intermediate product B15.

 Synthesis of the second intermediate C15 (Scheme 15): The first intermediate B15 (2.7 g, 9.2 mmol) was dissolved in THF (50 ml) then diethylene glycol anhydride (1.4 g, 12 mmol) and DMAP (1.5 g) , 12 mmol), heated to reflux for 12 hours. The obtained second intermediate C15 was used in the next step without further purification.

Synthesis of cytosolic derivative 15 (Scheme 15): cytarabine (1.3 g, 5 mmol), second intermediate C15 (2.1 g, 5 mmol), PyBOP (2.9 g, 5.5 mmol), and DMAP (0.06 g, 0.5 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 25/1) to give cytarabine derivative 15 (120.6 mg). The purity of LC (UV 254 nm) was 93%. LC-MS m/z 636 [M + H] + (M. C ₃₀ H ₄₁ N ₃ O ₁₂ , molecular weight 635). Synthesis 16:

Synthesis of first intermediate product B16 (Scheme 16): Mix salicylic acid (3.45 g, 25 mmol) with 1,12-dodecanediol (10.1 g, 50 mmol), then add 5 drops of concentrated sulfuric acid, heat After reacting at 80 ° C for 5 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) gave the first intermediate product B16

Synthesis of the second intermediate C16 (Scheme 16): The first intermediate B16 (0.966 g, 3 mmol) was dissolved in THF (30 ml) then diglycol (0.452 g, 3.9 mmol) and DMAP (0.475) g, 3.9 mmol), heated to reflux for 12 hours, and the obtained second intermediate C16 was used in the next step without further purification.

Synthesis of cytosolic derivative 16 (Scheme 16): cytarabine (0.729 g, 3 mmol), second intermediate C16 ( 1.314 g, 3 mmol), PyBOP ( 1.848 g, 3.3 mmol) and DMAP ( 0.06 g, 0.5 mmol) was dissolved in DMF (30 ml). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 15/1) to give cytarabine derivative 16 (26.6 mg). LC (UV 254 nm) purity 80%. LC-MS m/z 664 [M + H] + (Molecular formula C ₃₂ H ₄₅ N ₃ 0 ₁₂ , molecular weight 663 ).

Synthesis of the first intermediate product B17 (Scheme 17): Salicylic acid (13. 8 g, 100 mmol) was mixed with 1,6-hexanediol (23.6 g, 200 mmol), followed by the dropwise addition of 5 drops of sulphuric acid. The mixture was heated to 80 ° C for 5 hours, and after completion of the reaction, purification was carried out by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) to obtain the first intermediate product B17.

Synthesis of the second intermediate C17 (Scheme 17): The first intermediate B17 (1.5 g, 6.3 mmol) was dissolved in THF (30 ml) then phthalic anhydride (1.05 g, 6.3 mmol) and DMAP ( 0.84 g, 6.9 mmol), heated to reflux for 12 hours. The obtained second intermediate C17 was used in the next reaction without further purification.

Synthesis of cytosolic derivative 17 (Scheme 17): cytarabine (1.5 g, 6.3 mmol), second intermediate C17 (2.43 g, 6.3 mmol), PyBOP (3.28 g, 6.3 mmol) and DMAP (0.08 g, 0.65 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 30/1) to give cytarabine derivative 17 (66.1 mg). LC (UV 254 nm) purity 96%. LC-MS m/z 612 [M + H] + (M. C _{3 .} H ₃₃ N ₃ 0 s, molecular weight 611 ). synthetic route

Synthesis of the first intermediate product B18 (Scheme 18): Mix salicylic acid (6.9 g, 50 mmol) with 1,8-octanediol (14. 6 g, 100 mmol), then add 3 drops of concentrated sulfuric acid and heat to After reacting at 80 ° C for 5 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) afforded the first intermediate product B18.

Synthesis of second intermediate C18 (Scheme 18): The first intermediate B18 (2.000 g, 7.5 mmol) was dissolved in THF (100 mL) then phthalic anhydride (1.45 g, 9.8 mmol) and DMAP ( 1.20 g, 9.8 mmol), heated to reflux for 12 hours. The obtained second intermediate C18 was used in the next reaction without further purification.

Synthesis of cytosolic derivative 18 (Scheme 18): cytarabine (1.82 g, 7.5 mmol), second intermediate C18 (3.11 g, 7.5 mmol), PyBOP (4.3 g, 8.3 mmol) and DMAP (91.5 mg, 0.75 mmol) was dissolved in DMF (15 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 15/1) to give cytarabine derivative 18 (78.8 mg). LC (UV 254 nm) purity 99%. LC-MS m/z 640 [M + H] + (^ sub C ₃₂ H ₃₇ N ₃ 0 s, molecular weight 639 ).

Synthesis of first intermediate product B19 (Scheme 19): Salicylic acid (13.8 g, 100 mmol) was mixed with 1,10-decanediol (34.8 g, 200 mmol), then 5 drops of concentrated sulfuric acid were added dropwise and heated to After reacting at 80 ° C for 5 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) afforded the first intermediate product B19.

Synthesis of second intermediate C19 (Scheme 19): The first intermediate B19 (3.0 g, 10 mmol) was dissolved in THF (100 ml) then phthalic anhydride (1.8 g, 12 mmol) and DMAP (1.5) g, 12 mmol), heated to reflux for 12 hours. The obtained second intermediate C19 was used in the next step without further purification.

Synthesis of cyanosine derivative 19 (Scheme 19): cytarabine (2.4 g, 10 mmol), second intermediate C19 (4.4 g, 10 mmol), PyBOP (5.7 g, 11 mmol), and DMAP (0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 25/1) to give cytarabine derivative 19 (48 mg). The purity of LC (UV 254 nm) was 94%. LC-MS m/z 668 [M + H] + (M. C ₃₄ H ₄₁ N ₃ O _n , molecular weight 667).

Synthesis of first intermediate product B20 (Scheme 20): Mix salicylic acid ( 1.38 g, 10 mmol) with 1,12-dodecanediol (4.04 g, 20 mmol), then add 3 drops of concentrated sulfuric acid, heat The reaction was carried out at 80 ° C for 5 hours, and after completion of the reaction, purification was carried out by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) to obtain a first intermediate product B20.

Synthesis of second intermediate C20 (Scheme 20): The first intermediate B20 ( 1.96 g, 6.1 mmol) was dissolved in THF (40 mL) then phthalic anhydride (0.9 g, 6.1 mmol) and DMAP ( 0.74 g, 6.1 mmol), heated under reflux for 12 hours, and the second intermediate C20 obtained was used in the next step without further purification.

Synthesis of cyanosine derivative 20 (Scheme 20): cytarabine (1 g, 4.1 mmol), second intermediate C20 ( 2.76 g, 6.1 mmol), PyBOP ( 2.35 g, 4.5 mmol ) and DMAP (0.05 g, 0.41 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 15/1) to give cytarabine derivative 20 (18.1 mg). LC (UV 254 nm) purity 96%. LC-MS m/z 696 [M + H] + (Molecular formula C^H NsOn, molecular weight 695). Synthetic route 2

Synthesis of first intermediate product B21 (Scheme 21): Salicylic acid (13. 8 g, 100 mmol) was mixed with 1,6-hexanediol (23.6 g, 200 mmol), then 5 drops of concentrated sulfuric acid were added dropwise and heated to After reacting at 80 ° C for 5 hours, after completion of the reaction, purification was carried out by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) to obtain a first intermediate product B21.

Synthesis of the second intermediate C21 (Scheme 21): The first intermediate B21 (7.2 g, 30 mmol) was dissolved in THF (20 ml), then anhydride (5.6 g, 36 mmol) and DMAP (4.4 g, 36) Mmmol), heated to reflux for 12 hours. The obtained second intermediate C21 was used in the next step without further purification.

Synthesis of cytosolic derivative 21 (Scheme 21): cytarabine (2.4 g, 10 mmol), second intermediate C21 (3.9 g, 10 mmol), PyBOP (5.7 g, 11 mmol), and DMAP (0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 10/1) gave cytarabine derivative 21 (151.3 mg). LC (UV 254 nm) purity 96%. LC-MS m/z 618 [M + H] + (Molecular formula CsoH^On, molecular weight 617). Synthetic route 22:

Synthesis of first intermediate product B22 (Scheme 22): Mix salicylic acid (6.9 g, 50 mmol) with 1,8-octanediol (14. 6 g, 50 mmol), then add 5 drops of concentrated sulfuric acid and heat to After reacting at 80 ° C for 5 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) afforded the intermediate intermediate B22.

Synthesis of the second intermediate C22 (Scheme 22): The first intermediate B22 (1.4 g, 5.3 mmol) was dissolved in THF (20 ml), then anhydride (0.97 g, 6.3 mmol) and DMAP (0.77 g, 6.3) Mmmol), heated to reflux for 12 hours. The obtained second intermediate C22 was used in the next step without further purification.

Synthesis of cytosolic derivative 22 (Scheme 22): cytarabine (1.7 g, 6.9 mmol), second intermediate C22 (2.2 g, 5.3 mmol), PyBOP (3.6 g, 6.9 mmol), and DMAP (0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 25/1) to give cytarabine derivative 22 (129.2 mg). LC (UV 254 nm) purity 98%. LC-MS m/z 646 [M + H] + (M.s. CH NsOn, molecular weight 645 ). Synthetic route 23:

Synthesis of the first intermediate product B ² 3 (Scheme ² 3): Salicylic acid (13.8 g, 100 mmol) was mixed with 1,10-decanediol (34.8 g, 200 mmol), and then 5 drops of concentrated sulfuric acid were added dropwise. After heating to 80 ° C for 12 hours, after completion of the reaction, purification by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) afforded the first intermediate product B23.

Synthesis of the second intermediate C23 (Scheme 23): The first intermediate B23 (3.0 g, 10 mmol) was dissolved in THF (100 ml), then anhydride (2.0 g, 13 mmol) and DMAP (1.6 g, 13) Mmmol), heated to reflux for 12 hours. The obtained second intermediate C23 was used in the next step without further purification.

分子量 673 ) 。 Synthesis of cyanosine derivative 23 (Scheme 23): cytarabine (3.2 g, 13 mmol), second intermediate C23 (4.5 g, 10 mmol), PyBOP (7.8 g, 15 mmol), and DMAP (0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 25/1) to give cytarabine derivative 23 (98.3 mg). LC (UV 254 nm) purity 91%. LC-MS m/z 674 [M + H]+ (Molecular Formula

Molecular weight 673).

Synthesis of first intermediate product B24 (Scheme 24): Mix salicylic acid ( 1.38 g, 10 mmol) with 1,12-dodecanediol (4.04 g, 20 mmol), then add 3 drops of concentrated sulfuric acid, heat After reacting at 80 ° C for 5 hours, after completion of the reaction, purification was carried out by column chromatography (silica gel, developing solvent: petroleum ether / ethyl acetate = 5 / 1) to obtain a first intermediate product B24.

Synthesis of the second intermediate C24 (Scheme 24): The first intermediate B24 (0.8 g, 2.5 mmol) was dissolved in THF (30 ml), then anhydride (0.5 g, 3.2 mmol) and DMAP (0.4 g, 3.2). Mmmol), heated to reflux for 12 hours. The obtained second intermediate C24 was used in the next step without further purification.

Synthesis of cytosolic derivative 24 (Scheme 24): cytarabine (0.9 g, 3.9 mmol), second intermediate C24 (1.5 g, 3.2 mmol), PyBOP (2.0 g, 3.9 mmol), and DMAP (0.012 g, 0.1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 25/1) to give cytarabine derivative 24 (26.6 mg). LC (UV 254 nm) purity 84%. LC-MS m/z 702 [M + H] + (Molecular formula C ₃₆ H ₅₁ N ₃ 0 s, molecular weight 701 ). Synthetic route 25:

'N0 ₂

Dish ₂

 M25

 H

第十中间产物 M25的合成（路线 25 ) ： 将 1,8-辛二酸（3.48 g, 20 mmol )加 入到 SOCl₂ ( 15 ml )中， 加 DMF ( 1滴），加热回流，反应 3小时。旋去 SOCl₂, 得到的第十中间产物 M25不用进一步纯化， 直接用于下一步反应。

Synthesis of the tenth intermediate product M25 (Scheme 25): Add 1,8-suberic acid (3.48 g, 20 mmol) to SOCl ₂ (15 ml), add DMF (1 drop), heat to reflux, and react for 3 hours. . The SOCI ₂ was unscrewed and the obtained intermediate product M25 was used in the next step without further purification.

Synthesis of the eleventh intermediate product N25 (Scheme 25): o-Nitroaniline ( 2.76 g, 20 mmol) was dissolved in benzene (20 ml) and then pyridine (8 ml) was added, and o-nitroaniline was added dropwise at room temperature. Dissolved in benzene (40 ml), the tenth intermediate product M25 (4.04 g, 20 mmol). The reactant was dissolved in water, the pH was adjusted to be acidic, the precipitate was filtered, washed with water, the precipitate was dissolved in isopropyl alcohol, and recrystallized to precipitate the eleventh intermediate N25 of the smectite solid.

Synthesis of cytosolic derivative 25 (Scheme 25): cytarabine (0.42 g, 1.7 mmol), eleventh intermediate Ν25 (0.5 g, 1.7 mmol), PyBOP (0.97 g, 1.87 mmol) and DMAP (0.021 g, 0.17 mmol) was dissolved in DMF (5 mL). The reaction solution was poured into water to precipitate a solid, and the solid was purified by column chromatography (silica gel, solvent: methylene chloride / decyl alcohol = 15/1) to obtain cytarabine derivative 25 (31.3 mg). LC (UV 254 nm) purity 95%. LC-MS m/z 520 [M + H] ⁺ (Molecular formula C ₂₃ H ₂₉ N ₅ 0 ₉ , molecular weight 519 ). Synthetic route 26:

Synthesis of the twelfth intermediate product S26 (Scheme 26): o-Nitrobenzoic acid (3.34 g, 20 mmol) and SOCl ₂ (12 ml) were dissolved in DMF (10 ml) and refluxed for 2 hours. The twelfth intermediate product S26 obtained was used in the next step without further purification.

Synthesis of the thirteenth intermediate product Τ26 (Scheme 26): The twelfth intermediate product S26 (3.6 g, 20 mmol) was dissolved in THF (20 ml) and added dropwise to aqueous solution of alanine (1.8 g, 20 mmol) In the middle, a sodium hydroxide (0.8 g, 20 mmol) aqueous solution was added dropwise to control the pH = 8-9, and the control temperature was about 10 °C. After 30 min, the addition was completed and stirred for 40 min. The mixture was adjusted to pH = 3, THF was evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated

Synthesis of the fourteenth intermediate U26 (Scheme 26): cytarabine (1.56 g, 6 mmol), thirteenth intermediate Τ26 ( 1.43 g, 6 mmol), PyBOP (3.12 g, 6 mmol) and DMAP (0.07 g, 0.6 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 6/1) to obtain the fourteenth intermediate product U26.

Synthesis of cytarabine derivative 26 (Scheme 26): The fourteenth intermediate U26 (about 40 mg, 0.086 mmol) was dissolved in THF (30 ml), the catalytic amount of Pd/C was added, hydrogen was introduced, and the mixture was stirred for 2 h. , filtration, and the filtrate was spin-dried to obtain cytarabine derivative 26 (24.7 mg). LC (UV 254 nm) purity 94%. LC-MS m/z 434 [M + H] ⁺ (M. C ₁₉ H ₂₃ N ₅ 0 ₇ , molecular weight 433).

Synthesis of the twelfth intermediate product S27 (Scheme 27): o-Nitrobenzoic acid (1.67 g, 10 mmol) and S0C1 ₂ (10 ml) were dissolved in DMF (10 ml) and refluxed for 2 hours. The twelve intermediate product S27 was used directly in the next reaction without further purification.

 Synthesis of the thirteenth intermediate product Τ27 (Scheme 27): The twelfth intermediate product S27 (1.8 g, 10 mmol) was dissolved in THF (20 ml) and added dropwise to an aqueous solution of aminohexanoic acid (1.5 g, 12 mmol) In the middle, a sodium hydroxide (0.96 g, 24 mmol) aqueous solution was added dropwise to control the pH = 8-9, and the control temperature was about 10 °C. After 30 min, the mixture was stirred for 40 min. The pH was adjusted to pH = 3, THF was evaporated under reduced pressure, and the aqueous mixture was evaporated to ethyl acetate. EtOAc was evaporated. Alkane/nonanol = 10/1) gave the thirteenth intermediate T27.

 Synthesis of the fourteenth intermediate U27 (Scheme 27): cytarabine (2.4 g, 10 mmol), thirteenth intermediate Τ27 (2.8 g, 10 mmol), PyBOP (5.7 g, 11 mmol) and DMAP (0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 10/1) to obtain the fourteenth intermediate product U27.

Synthesis of cytarabine derivative 27 (Scheme 27): The fourteenth intermediate U27 (about 40 mg, 0.08 mmol) was dissolved in THF (30 ml), a catalytic amount of Pd/C was added, hydrogen gas was added thereto, stirred for 2 hours, filtered, and the filtrate was dried to give cytarabine derivative 27 (36.2 mg). LC (UV 254 nm) purity 97%. LC-MS m/z 476 [M + H] ⁺ (Molecular formula C ₂₂ H ₂₉ N ₅ 0 ₇ , molecular weight 475 ).

第十五中间产物 V28的合成（路线 28)： 将邻硝基苯曱酸（ 1.67 g, 10 mmol) , 氨基己醇（1.17 g, 10 mmol) , PyBOP (5.72g, 10 mmol )和 DMAP ( 0.12 g, 1 mmol )溶于 DMF (10 ml) , 室温搅拌 12小时。 反应液倒入水中， 乙酸乙酯 萃取三次， 乙酸乙酯液用无水硫酸钠干燥， 过滤， 滤液减压蒸干， 再通过柱层 析色语提纯（硅胶，展开剂： 二氯曱烷 /曱醇 =50/1),得到第十五中间产物 V28。 第十六中间产物 W28的合成（路线 28 ):将第十五中间产物 V28( 2.1 g, 8 mmol ) 溶于 40毫升四氢呋喃中， 然后加入邻苯二曱酸酐（ 1.48 g, 10 mmol)和 DMAP ( 1.22 g, 10 mmol) , 加热回流反应 12小时。 得到的第十六中间产物 W28不 经进一步纯化， 直接用于下一步反应。

Synthesis of the fifteenth intermediate V28 (Scheme 28): o-Nitrobenzoic acid (1.67 g, 10 mmol), aminohexanol (1.17 g, 10 mmol), PyBOP (5.72 g, 10 mmol) and DMAP ( 0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water and extracted with EtOAc EtOAc EtOAc EtOAc. The decyl alcohol = 50/1) gave the fifteenth intermediate product V28. Synthesis of the sixteenth intermediate W28 (Scheme 28): The fifteenth intermediate V28 (2.1 g, 8 mmol) was dissolved in 40 mL of tetrahydrofuran, then phthalic anhydride (1.48 g, 10 mmol) and DMAP were added. ( 1.22 g, 10 mmol), heated to reflux for 12 hours. The obtained sixteenth intermediate W28 was used in the next step without further purification.

Synthesis of the seventeenth intermediate Υ28 (Scheme 28): cytarabine (lg, 4.1 mmol), the sixteenth intermediate W28 ( 1.86 g, 4.5 mmol), PyBOP ( 2.35 g, 4.5 mmol) and DMAP ( 0.05 g, 0.4 mmol) was dissolved in DMF (10 ml) and stirred at room temperature for 12 h. The reaction solution is poured into the water. The mixture was extracted with EtOAc (EtOAc) EtOAc. , the seventeenth intermediate product Y28 is obtained.

Synthesis of cytarabine derivative 28 (Scheme 28): The seventeenth intermediate product Υ28 (50 mg, O.lmmol) was dissolved in THF (30 ml), a catalytic amount of Pd/C was added, hydrogen was introduced, and the mixture was stirred for 2 h. After filtration, the filtrate was spun dry to give cytarabine derivative 28 (45.1 mg). The purity of LC (UV 254 nm) was 94%. LC-MS m / z 610 [ M + H] + ( molecular formula _{C 3 .H 35 N 5 0 9} , 609 molecular weight). Synthetic route 29:

Synthesis of the fifteenth intermediate V29 (Scheme 29): Niacin (1.35 g, 11 mmol), 6-amino-1-hexanol B (1.17 g, 10 mmol), PyBOP (5.72 g, 11 mmol) and DMAP (0.12 g, 1 mmol) was dissolved in DMF (10 mL). The reaction mixture was poured into water and extracted with EtOAc EtOAc EtOAc EtOAc EtOAc. The decyl alcohol = 15/1) gave the fifteenth intermediate product V29. Synthesis of the sixteenth intermediate W29 (Scheme 29): The fifteenth intermediate V29 (1.14 g, 5.1 mmol) was dissolved in 40 mL of tetrahydrofuran, then phthalic anhydride (0.83 g, 5.6 mmol) and DMAP were added. (0.68 g, 5.6 mmol), heated to reflux for 12 hours. The obtained sixteenth intermediate W29 was used in the next step without further purification. Synthesis of cyanosine derivative 29 (Scheme 29): cytarabine (1 g, 4.1 mmol), sixteenth intermediate W29 ( 1.89 g, 5.1 mmol), PyBOP ( 2.35 g, 4.5 mmol ) and DMAP (0.05 g, 0.41 mmol) was dissolved in DMF (10 ml) and stirred at room temperature for 12 h. The reaction mixture was poured into water, and the mixture was evaporated. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj Alcohol = 15/1) to give cytarabine derivative 29 (32.1 mg). The purity of LC (UV 254 nm) was 92%. LC-MS m/z 596 [M + H] + (M. C ₂₉ H ₃₃ N ₅ 0 ₉ , molecular weight 595 ).

Synthetic drug tumor cytotoxicity test protocol

 Inhibition of tumor cells by the cytarabine derivative of the invention and preparation thereof

 Test material

 1) Cell line:

The HL-60 cell line was suspended and grown in RPMI 1640 cell culture medium containing 10% fetal bovine serum (Hyclone). The normal culture was maintained at an initial cell concentration of about 3*10 ⁵ /ml, and once every three days for 1:3 passage. . Passage (5* 10 ⁵ /ml) one day before the experiment, the cell concentration during the experiment

Between 7.5 and 10*10 ⁵ /ml.

The BEL-7402 cell line and the HT-29 cell line were adherently grown and cultured in D-MEM cell culture medium of 10% fetal bovine serum (Hyclone). The initial cell concentration of the conventional culture was about 3*10 ⁵ /ml, 2 ~3 days 1: 3 pass once. On the day before the experiment, 1: 2 passage, the cell concentration during the experiment was between 5~10*10 ⁵ /ml.

 2) Dissolution and dilution of the drug: According to the weight and molecular weight of the cytarabine derivative provided, first add DMSO 100~200 μΐ, then add physiological saline (NS), so that the concentration of the drug obtained after dilution is 5 mM (note The final concentration of DMSO does not exceed 10%).

 3) D-MEM or RPMI 1640 Cell Culture Medium, Gibco

 4) fetal bovine serum, Hyclone

5) Cell digestive juice, 0.25 % Trypsin + 0.02 % EDTA 6) PBS phosphate buffer

 7) MTT solution, MTT dry powder (Sigma), fully dissolved in PBS, formulated into 5 mg/ml, 0.22 μηι microporous membrane, filtered, and stored at -20 °C.

 8) 10% acidification SDS, 0.01N HC1

 9) Centrifuge tubes, straws, etc. (BD company), 96-well plate (Costar)

 2. Steps:

1) Cell seeding: Cells grown 24 hours after passage were in good growth state. The cells were routinely harvested, and the cell concentration was adjusted to 2 x ^{10 5} /ml (adherent cells) ~ 3 x ^{10 5} /ml (suspended cells) with fresh medium.

The adherent cells were inoculated with 100 μΐ/well, cultured in a 37 ° C, 5% CO ₂ incubator for 24 h, and the old culture solution was discarded, and fresh culture solution was added at 95 μM/well.

 Suspension cells were inoculated directly at 95 μΐ/well.

2) Drug treatment: There are 6 concentration gradients for each drug, 3 duplicate wells for each concentration, and 5 duplicate wells for the drug blank control group. Ara-C control was performed at the same time for each test. The concentrations of HT-29 and BEL-7402 cells were 5, 2.5, 1.25, 0.625, 0.3125, 0.16 mM, 5 μl per well, and the final concentrations were 0.25, 0.125, 0.0625, 0.03125, 0.016, 0.008 mM, respectively. Add 5 μΐ saline to the group; the concentration of HL60 cells added to the drug is 5χ10 ^-3 , 2.5X10" ³ , 1.25X10" ³ , 0·625χ10 ^-3 , 0·3125χ10 ^-3 , 0.16xlO" ³ mM , corresponding to the final concentration The order was 2.5χ1 (Τ ⁴ , 1.25χ1 (Τ ⁴ , 6.25χ1 (Τ ⁵ , 3·125χ10 ^-5 , 1·6χ10 ^-5 , 8xlO” ⁶ mM, and the control group was added with 5 μΐ of normal saline.

3) Cell culture and detection: After adding the drug, 37. C, 5% C0 ₂ incubator for 72h, then add MTT 10 μΐ per well, continue to culture for 4 h, add 100 μΐ 10% SDS (containing 0.01 N HC1 ) per well, and use Bio-rad 680 after 24 h. The absorbance (A) of each well was measured by an ELISA reader with a detection wavelength of 570 nm and a reference wavelength of 630 nm.

4) Calculation: First, average the absorbance of each replicate well (remove the data from over-disparity), calculate the inhibition rate (IR) for each drug concentration of each cell, IR(%)=(1- A _n /A ₀ )xl00 %, The average absorbance of the experimental wells, Ao is the average absorbance of the drug blanks. Using the EXCEL software, plot the drug concentration effect curve and select a reasonable calculation method to calculate the drug concentration (IC ₅₀ ) for 50% cell survival.

 Fig. 3 is a graph showing the drug concentration-inhibition rate of the cytosine derivative of the present invention inhibiting the BEL-7402 liver cancer cell line. JF007, JF017, JF019, JF020 and JF029 are cytarabine derivatives synthesized according to the synthetic routes 7, 17, 19, 20 and 29, respectively.

Table 1 lists the representative cytarabine derivatives inhibiting the biological activity of different tumor cells. Among them, JF002, JF003, JF004 and JF006 are cytarabine derivatives synthesized according to synthetic routes 2, 3, 4 and 6, respectively.

Table 1

试验证明， 本发明的阿糖胞苷衍生物具有抗实体瘤的生物活性。

Tests have shown that the cytarabine derivatives of the present invention have biological activity against solid tumors.

Claims

Claim A cytarabine derivative characterized in that the cytarabine derivative is a compound having the following formula (I):

General formula -I Wherein X is any one of OH, 0-P(0)(OR) ₂ and a phosphate group, and the phosphate group includes a monophosphate group, a diphosphorus group, and a triphosphate group; Is any one of 11, C _M6 alkyl, cycloalkyl, benzyl, phenyl and aromatic ring groups;  Wherein, A is the second drug-based or functional group in the multi-target drug, and any one of the following formulas is used.

^ is 11, an alkane group, an alkenyl group, an alkynyl group, and an aryl group; R ² is any one of a nitro group, an amino group, a substituted amino group, a halogen atom group, a nitrile group, and an amide group;  L is a connection unit and is represented by any of the following structural units:

Wherein, 11 = 0, 1-5; m = 0, 1-18; the curve ", /" indicates that the above-mentioned linking unit L is covalently bonded to its adjacent group; the benzene ring or ring in the linking unit L The two groups on the alkane ring are ortho, meta or para.  The cytarabine derivative according to claim 1, wherein the cytarabine derivative of X = OH in the formula (I). The cytarabine derivative according to claim 1 or 2, wherein the structural formula of the cytarabine derivative is any one of the following structural formulae:

51

The synthetic route of cytarabine derivatives includes the following steps: (1) mixing salicylic acid with an aliphatic diol, then adding 3 to 5 drops of concentrated sulfuric acid, heating to 80 ° C ~ 12 hours, after the reaction is finished, evaporating under reduced pressure or by column chromatography Purification, get the first Interproduct (B); (2) Dissolving the first intermediate product (B) of the step (1) in CH ₂ C1 ₂ or tetrahydrofuran, then adding the acid anhydride compound and DMAP, heating the reflux reaction for 5-12 hours or stirring at room temperature for 24 hours, and obtaining the first The intermediate product (C) was used in the next step without further purification;  (3) Dissolving cytarabine, second intermediate product (C), PyBOP and DMAP in DMF, stirring at room temperature for 12-24 hours, and directly purifying the reaction solution by column chromatography to obtain the general formula (I). The cytidine derivative or the reaction solution is poured into water, and extracted with ethyl acetate three times. The ethyl acetate solution is dried over anhydrous sodium sulfate and filtered, and the filtrate is evaporated to dryness, and purified by column chromatography. A cytarabine derivative of the formula (I).  The synthetic route of the cytarabine derivative according to claim 4, wherein the aliphatic diol is ethylene glycol, butylene glycol, 1,6-hexanediol, 1, 8 - any one of octanediol, 1,10-nonanediol, and 1,12-dodecanediol; the anhydride compound is succinic anhydride, phthalic anhydride, glutaric anhydride, and diglycolic anhydride Any of them.  6. The synthetic route of cytarabine derivatives, including the following steps: (1) Salicylic acid and sodium carbonate are dissolved in 3-chloropropanol, heated under reflux for 2.5 hours, and the obtained reaction solution is poured into cold water to separate the layers, and the organic layer is washed with NaHC0 ₃ solution to weakly alkaline, and the organic layer is reused. After washing with a small amount of saturated brine, it is dried over anhydrous sodium sulfate, filtered, and the filtrate is evaporated to dryness to give a third intermediate (D) which is directly used for the next reaction; (2) Dissolving the third intermediate product (D) in CH ₂ C1 ₂ or tetrahydrofuran, then adding the acid anhydride compound and DMAP, stirring the reaction at room temperature for 24 hours, filtering the reaction liquid, and evaporating the filtrate under reduced pressure to obtain a fourth intermediate. The product (E) is used directly in the next reaction; (3) Dissolving cytarabine, fourth intermediate (E), PyBOP and DMAP in DMF, stirring at room temperature for 12-24 hours, and directly purifying the reaction liquid by column chromatography to obtain arabinose of general formula (I) The cytidine derivative or the reaction solution is poured into water, and extracted with ethyl acetate three times. The ethyl acetate solution is dried over anhydrous sodium sulfate and filtered, and the filtrate is evaporated to dryness under reduced pressure and purified by column chromatography. A cytarabine derivative of formula (I). The synthetic route of the cytarabine derivative according to claim 6, wherein the acid anhydride compound is any one of succinic anhydride, phthalic anhydride, glutaric anhydride, and diglycolic anhydride. Kind.  8. The synthetic route of cytarabine derivatives, including the following steps:  (1) The decyl salicylate and aminopropanol are dissolved in dioxane, stirred at room temperature for 12 hours, the solvent is removed by rotary evaporation, saturated brine is added, and the pH is adjusted to 4-5 with hydrochloric acid, and then extracted with ethyl acetate. Then, the ethyl acetate solution was dried over anhydrous sodium sulfate, filtered, and the filtrate evaporated to dryness under reduced pressure, and the obtained intermediate product (F) was directly used for the next reaction; (2) Dissolving the fifth intermediate product (F) in CH ₂ C1 ₂ or tetrahydrofuran, then adding the acid anhydride compound and DMAP, stirring the reaction at room temperature for 24 hours, filtering the reaction liquid, and evaporating the filtrate under reduced pressure to obtain a sixth intermediate portion. The product (G) was used in the next step without further purification;  (3) Dissolving cytarabine, sixth intermediate (G), PyBOP and DMAP in DMF, stirring at room temperature for 12-24 hours, and purifying the reaction directly by column chromatography to obtain arabin of formula (I) The cytidine derivative or the reaction solution is poured into water, and extracted with ethyl acetate three times. The ethyl acetate solution is dried over anhydrous sodium sulfate and filtered, and the filtrate is evaporated to dryness. (I) cytarabine mites.  The synthetic route of the cytarabine derivative according to claim 8, wherein the acid anhydride compound is any one of succinic anhydride, phthalic anhydride, glutaric anhydride, and diglycolic anhydride. Kind.  10. The synthetic route of cytarabine derivatives, including the following steps: (1) o-decyloxybenzoic acid and SOC2 _{2 are} dissolved in dichloromethane, heated under reflux for 4 hours, and the seventh intermediate product (H) obtained by removing the solvent and excess SOCl ₂ is removed by rotary evaporation;  (2) Dissolving aminopropanol in dichlorosilane, and cooling to 0 °C, adding the seventh intermediate product (H) dissolved in dichloromethane, adding 0.5 hours, and then stirring at room temperature for 5 hours. The solution was acidified to pH = 5, and the solvent was evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated, evaporated. Solvent, the obtained eighth intermediate product (J) is directly used in the next reaction without purification; (3) dissolving the eighth intermediate product (J) in tetrahydrofuran, then adding an acid anhydride compound and DMAP, The reaction was stirred at room temperature for 24 hours, filtered, and the filtrate was evaporated to dryness, and the obtained ninth intermediate product (K) was directly used for the next reaction without purification;  (4) Dissolving cytarabine, ninth intermediate product (K), PyBOP and DMAP in DMF, stirring at room temperature for 12 to 24 hours, and directly purifying the product by column chromatography to obtain a product or pouring the reaction solution into water. The mixture was extracted three times with ethyl acetate. The ethyl acetate solution was dried over anhydrous sodium sulfate and filtered, and the filtrate was evaporated to dryness, and purified by column chromatography to give the cytosine derivative of formula (I).  The synthetic route of the cytarabine derivative according to claim 10, wherein the acid anhydride compound is any one of succinic anhydride, phthalic anhydride, glutaric anhydride, and diglycolic anhydride. Kind.  12. The synthetic route of cytarabine derivatives, including the following steps: (1) Adding an aliphatic dibasic acid to S0C1 ₂ , adding DMF, heating under reflux, and reacting for 3 hours, and spinning off S0C1 ₂ , the obtained intermediate product (M) is directly used for the next reaction without further purification;  (2) Dissolving o-nitroaniline in benzene and adding pyridine, adding dropwise to the tenth intermediate product (M) dissolved in benzene, reacting for 2 to 4 hours, evaporating the solvent, dissolving the reactant in water, adjusting pH The value is acidic, the precipitate is filtered, washed with water, the precipitate is dissolved in isopropanol, recrystallized, and the eleventh intermediate (N) of the solid is precipitated;  (3) Dissolving cytarabine, the eleventh intermediate product (N), PyBOP and DMAP of the solid in DMF, stirring at room temperature for 12 hours, pouring the reaction solution into water, precipitating the solid, and purifying by column chromatography. A cytarabine derivative of the formula (I) is obtained.  The synthetic route of the cytarabine derivative according to claim 12, wherein the aliphatic dibasic acid is oxalic acid, succinic acid, 1,6-hexanediol, and 1,8- Any of suberic acid.  14. The synthetic route of cytarabine derivatives, including the following steps: (1) o-Nitrobenzoic acid and SOC2 _{2 were} dissolved in DMF, and refluxed for 2 hours, and the twelfth intermediate product (S) obtained was used in the next step without further purification. (2) Dissolving the twelfth intermediate product (S) in tetrahydrofuran, adding dropwise to the aqueous solution of the amino acid compound, and simultaneously adding a sodium hydroxide aqueous solution to control the pH=8~9, and controlling the temperature at about 10 °C, 30 minutes After the completion of the dropwise addition, stirring for 40 minutes and adjusting the pH to 3, the tetrahydrofuran was evaporated under reduced pressure, the aqueous solution was extracted with EtOAc EtOAc EtOAc EtOAc. Thirteenth intermediate product (T);  The amino acid compound is alanine or aminocaproic acid;  (3) Dissolving cytarabine, thirteenth intermediate (T), PyBOP and DMAP in DMF, stirring at room temperature for 12 hours, pouring the reaction solution into water, extracting three times with ethyl acetate, and using anhydrous sulfuric acid for ethyl acetate solution. The sodium is dried, filtered, and the filtrate is evaporated to dryness under reduced pressure, and then purified by column chromatography to obtain the fourteenth intermediate product (U);  (4) The fourteenth intermediate product (U) is dissolved in tetrahydrofuran, catalyzed by hydrogenolysis, stirred, filtered, and the filtrate is spin-dried to obtain a highly pure cytarabine derivative of the formula (I).  15. The synthetic route of cytarabine derivatives, including the following steps:  (1) Dissolving nitrobenzoic acid or nicotinic acid, hexanoicol, PyBOP and DMAP in DMF, stirring at room temperature for 12 hours, pouring the reaction solution into water, extracting three times with ethyl acetate, and using anhydrous sodium sulfate for ethyl acetate. Drying, filtering, and evaporating the filtrate under reduced pressure, and purifying by column chromatography to obtain the fifteenth intermediate product (V);  (2) dissolving the fifteenth intermediate product (V) in tetrahydrofuran, and then adding phthalic anhydride and DMAP, heating under reflux for 12 hours, the obtained sixteenth intermediate (W) was directly used for the next reaction without further purification;  (3) Dissolving cytarabine, the sixteenth intermediate product (W), PyBOP and DMAP in DMF, stirring at room temperature for 12 hours, pouring the reaction solution into water, extracting three times with ethyl acetate, and using anhydrous sulfuric acid for ethyl acetate solution. The sodium is dried, filtered, and the filtrate is evaporated to dryness under reduced pressure, and then purified by column chromatography to obtain the 17th intermediate product (Y);  (4) The seventeenth intermediate product (Y) is dissolved in tetrahydrofuran, catalyzed by hydrogenolysis, stirred, filtered, and the filtrate is spin-dried to obtain a highly purified cytarabine derivative of the formula (I).  16. A method for preparing a cytarabine derivative preparation, characterized in that: (1) dissolving the cytarabine derivative of the formula (I) into water, physiological saline, aqueous cyclodextrin solution, water-soluble organic solvent, nonionic surfactant, water-soluble lipid, fatty acid, Fatty acid Preparing a preparation solution by combining a solvent of any one or more of an ester and a phospholipid;  (2) The preparation solution is further diluted with physiological saline or glucose injection to prepare a cytarabine derivative.  The process for producing a cytarabine derivative preparation according to claim 16, wherein the organic solvent is ethanol, propylene glycol, glycerin, glycerin, polyethylene glycol, hydrazine, fluorenyl-difluorenyl hydrazine a combination solvent of any one or more of an amide and a dimercaptosulfoxide.  A cytarabine derivative preparation, which is characterized by the preparation of the preparation method of the cytarabine derivative preparation of claim 16.  The use of the cytarabine derivative of claim 1 for anticancer and antitumor.  The use of the cytarabine derivative according to claim 19 for anticancer and antitumor, characterized in that the cancer comprises leukemia, solid tumor, lung cancer, colon cancer, liver cancer, central nervous system tumor, ovarian cancer, renal cancer .  21. Use of a cytarabine derivative preparation according to claim 18 for anti-cancer and anti-tumor.  The use of the cytarabine derivative preparation according to claim 21 for anticancer and antitumor, characterized in that the cancer comprises leukemia, solid tumor, lung cancer, colon cancer, liver cancer, central nervous system tumor, ovarian cancer and kidney cancer.  The use of the cytarabine derivative preparation of claim 21 for anticancer and antitumor, characterized in that: the cytarabine derivative preparation is administered in combination with other chemotherapeutic drugs, and the other chemotherapeutic drugs include alkylation Agents, plant alkaloids, antibacterial and anti-tumor sulfonamides, platinum drugs, anti-metabolites and other known anti-cancer drugs.  The use of the cytarabine derivative preparation according to claim 23 for anticancer and antitumor, characterized in that, in the combined use, the use of at least one cytarabine derivative having the structural formula of claim 5 is used. Things.