WO2020114455A1 - 含有哌嗪酮的喹唑啉二酮盐类化合物、其制备方法、药物组合物和用途 - Google Patents

含有哌嗪酮的喹唑啉二酮盐类化合物、其制备方法、药物组合物和用途 Download PDF

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WO2020114455A1
WO2020114455A1 PCT/CN2019/123295 CN2019123295W WO2020114455A1 WO 2020114455 A1 WO2020114455 A1 WO 2020114455A1 CN 2019123295 W CN2019123295 W CN 2019123295W WO 2020114455 A1 WO2020114455 A1 WO 2020114455A1
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substituted
unsubstituted
membered aromatic
independently selected
methyl
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French (fr)
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徐柏玲
陈晓光
周洁
季鸣
李燕
盛莉
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中国医学科学院药物研究所
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Priority to CN201980088459.2A priority Critical patent/CN113272295A/zh
Publication of WO2020114455A1 publication Critical patent/WO2020114455A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/10—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing aromatic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/517—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with carbocyclic ring systems, e.g. quinazoline, perimidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the invention relates to a quinazoline-2,4-dione salt compound containing piperazinone as a PARP1/2 inhibitor and a preparation method thereof, a pharmaceutical composition containing the compound, and the compound as a medicine, especially As an antitumor drug or as a tumor drug sensitizer in combination with antitumor drugs.
  • Poly ADP ribose polymerase (Poly ADP Ribose Polymerase, PARP) is widely present in eukaryotic cells and is a ribozyme capable of catalyzing poly ADP ribosylation. This enzyme was first reported by Chambon et al in 1963. It has important functions such as maintaining chromosomal structural integrity, participating in DNA replication and transcription, and maintaining gene stability [D'Amours D. Et. Biochemical Journal, 1999, 342, 249– 268]. The PARP family contains at least 17 subtypes, of which PARP-1 is the most widely studied. [AmeJ.C.et.al.Bioessays,2004,26,882–893].
  • chemotherapeutic drugs kill tumor cells by directly or indirectly damaging the DNA chain, inhibiting the activity of PARP-1 can effectively block the repair of damaged DNA, thereby producing a synergistic antitumor effect with chemotherapeutic drugs.
  • PARP-1 inhibitors are more sensitive to mutations in breast cancer susceptibility genes (BRCA-1, BRCA-2), and can be used alone to treat BRCA-mutated tumors [Bryant, HEetal. Nature 2005, 434, 913-917] .
  • PARP-2 and PARP-1 have the highest homology [Kutuzov, M. M. et al. Molecular Biology 48, 485-495]. In terms of biological function, it is similar to PARP-1, but also different. Like PARP-1, PARP-2 can also be activated by damaged DNA [Ame, J.C.et.al.J Biol Chem 1999,274,17860-8]. Through the observation of the PARP-2 gene knockout model, it was found that the sensitivity of mice to ionizing radiation and methylation reagents increased, and the instability of the genome increased. In addition, PARP-2 also has unique physiological functions. Down-regulation of the PARP-2 gene will prevent cardiomyocyte hypertrophy [Geng, B. et al.
  • This patent is designed to synthesize piperazinone-containing quinazoline dione salt compounds that act on the PARP-1/2 target. These compounds have better antitumor activity in vivo and oral administration than the unsalted prototype compounds. Bioavailability aims to provide a new and better material basis for the treatment of diseases related to PARP-1/2.
  • the technical problem solved by the present invention is to provide a piperazinone-containing quinazoline-2,4(1H,3H)-dione salt derivative represented by formula I, a preparation method thereof, a pharmaceutical composition, and a preparation method thereof Use of PARP-1/2 inhibitors and their potential drugs, in the preparation of anti-tumor drugs or anti-tumor drug sensitizers.
  • the present invention provides the following technical solutions:
  • R 1 , R 2 , R 3 , R 4 and R 5 are independently selected from H, F, Cl and Br;
  • R 6 is selected from the following atoms or groups or structural fragments:
  • the benzene ring, nitrogen-containing six-membered aromatic heterocyclic ring, and five-membered aromatic heterocyclic ring may be mono-substituted or multi-substi
  • M is independently selected from different alkali metals (lithium, sodium, potassium, cesium) or alkaline earth metal salts (calcium, magnesium, barium).
  • said R 5 is independently preferably from F.
  • preferred compounds of the present invention include, but are not limited to, compounds represented by the general formula I-A:
  • R 1 , R 2 , R 3 and R 4 are independently selected from H, F, Cl and Br;
  • R′ 6 is selected from the following atoms or groups or structural fragments:
  • M is independently selected from different alkali metals (lithium, sodium, potassium, cesium) or alkaline earth metal salts (calcium, magnesium, barium).
  • R 1 , R 2 , R 3 , and R 4 are preferably independently selected from H and F.
  • preferred compounds of the present invention include, but are not limited to, compounds represented by the general formula I-B:
  • R 1 , R 2 , R 3 and R 4 are independently selected from H, F, Cl and Br;
  • Ar is selected from the following atoms or groups or structural fragments, including
  • the benzene ring, nitrogen-containing six-membered aromatic heterocyclic ring, and five-membered aromatic heterocyclic ring may be mono-substituted or multi-sub
  • M is independently selected from different alkali metals (lithium, sodium, potassium, cesium) or alkaline earth metal salts (calcium, magnesium, barium).
  • R 1 , R 2 , R 3 , R 4 and R 5 are preferably independently selected from H and F.
  • preferred compounds include but are not limited to:
  • the second aspect of the technical solution of the present invention provides a method for preparing the compound described in the first aspect.
  • the technical solution adopted includes the following steps: different quinazoline diones substituted with R 1 -R 4 and 3-containing R 5 -substituted
  • the selective alkylation of methoxycarbonyl benzyl bromide under the action of HMDS, after hydrolysis, contains different 3-((2,4-dioxo-3,4- substituted by R 1 -R 5 Dihydroquinazoline-1(2H)-yl)methyl)benzoic acid is then condensed with a piperidone derivative containing R 6 to obtain 1-benzylquinazoline dione compounds, and finally with different bases An acid-base reaction occurs to obtain a base addition salt of 1-benzylquinazoline dione compounds.
  • the definitions of R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , and M described herein are the same as the definitions of the compounds described
  • Reagents and reaction conditions (a) urea, 140°C, 6h; (b) i) lithium hexamethyldisilazide (HMDS), concentrated sulfuric acid, toluene, reflux, 2h, ii) substituted 5-(bromomethyl Methyl)-2-fluorobenzoic acid methyl ester, 145°C, 3h, iii) methanol, hexane, 70°C, 30min; (c) LiOH, MeOH, H 2 O, THF, 55°C, 2h; (d) 2 -(7-Azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), 1-hydroxybenzotriazole (HOBT), diisopropyl Ethylamine (DIEA), DMF (N,N-dimethylformamide), overnight (overnight); (e) MOH/H 2 O/40°C;
  • HMDS
  • R 1 , R 2 , R 3 , R 4 , R 5 , R 6 and M described therein are the same as the definitions of the compounds described in the first aspect of the present invention.
  • the starting materials and intermediates in the above reaction are easy to obtain, and each step of the reaction can be easily synthesized according to the reported literature or by a person skilled in the art using conventional methods in organic synthesis.
  • the compound described in the general formula I may exist in the form of a solvate or an unsolvate, and different solvents may be used for crystallization to obtain different solvates.
  • a third aspect of the technical solution of the present invention is to provide a pharmaceutical composition comprising the compound described in the first aspect of the technical solution of the present invention and a common pharmaceutical carrier.
  • the composition includes at least one compound of the present invention and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition is selected from tablets, capsules, pills, injections, sustained-release preparations, controlled-release preparations, or various microparticle delivery systems.
  • the pharmaceutical composition can be prepared according to methods known in the art.
  • the compound of the present invention may be combined with one or more pharmaceutically acceptable solid or liquid excipients and/or adjuvants to make any dosage form suitable for human or animal use.
  • the content of the compound of the present invention in its pharmaceutical composition is usually 0.1 to 95% by weight.
  • the compound of the present invention or the pharmaceutical composition containing it can be administered in unit dosage form, and the route of administration can be intestinal or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and Respiratory tract, skin, vagina, rectum, etc.
  • the dosage form for administration may be a liquid dosage form, a solid dosage form or a semi-solid dosage form.
  • Liquid dosage form can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and double emulsion), suspension, injection (including water injection, powder injection and infusion), eye drops Agents, nasal drops, lotions, liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric tablets, buccal tablets, dispersible tablets, chewable tablets, effervescent tablets, oral disintegrating tablets), capsules ( Including hard capsules, soft capsules, enteric capsules), granules, powders, pellets, dropping pills, suppositories, films, patches, gas (powder) aerosols, sprays, etc.; semi-solid dosage forms can be ointments, Gels, pastes, etc.
  • the compound of the present invention can be made into ordinary preparations, sustained release preparations, controlled release preparations, targeted preparations and various microparticle delivery systems.
  • auxiliary materials used in the manufacture of tablets, capsules, and coating agents are conventional auxiliary agents, such as starch, gelatin, gum arabic, silica, polyethylene glycol, and liquid dosage forms.
  • the solvents used are, for example, water, ethanol, propylene glycol, and vegetable oil. Such as corn oil, peanut oil, olive oil and so on.
  • the preparations containing the compound of the present invention may also contain other auxiliary agents, such as surfactants, lubricants, disintegrating agents, preservatives, flavoring agents, coloring agents, and the like.
  • diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.
  • the wetting agent can be water, ethanol, isopropyl Propyl alcohol, etc.
  • the binder may be starch slurry, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic, gelatin slurry, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl alcohol Cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch,
  • the tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer tablets and multi-layer tablets.
  • the active ingredient compound of the present invention can be mixed with a diluent and a glidant, and the mixture is directly placed in a hard capsule or a soft capsule.
  • the active ingredient of the compound of the present invention can also be made into granules or pellets with diluent, binder, disintegrant, and then placed in hard or soft capsules.
  • Various diluents, binders, wetting agents, disintegrating agents, glidants used to prepare the tablets of the compounds of the present invention can also be used to prepare capsules of the compounds of the present invention.
  • water, ethanol, isopropanol, propylene glycol or a mixture thereof can be used as a solvent and an appropriate amount of solubilizer, co-solvent, pH regulator, and osmotic regulator commonly used in the art can be added.
  • the solubilizer or co-solvent can be poloxamer, lecithin, hydroxypropyl- ⁇ -cyclodextrin, etc.
  • the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.
  • the osmotic regulator can It is sodium chloride, mannitol, glucose, phosphate, acetate, etc.
  • mannitol, glucose, etc. can also be added as proppants.
  • coloring agents may also be added to the pharmaceutical preparations.
  • preservatives may also be added to the pharmaceutical preparations.
  • fragrances may also be added to the pharmaceutical preparations.
  • flavoring agents may also be added to the pharmaceutical preparations.
  • the drug or pharmaceutical composition of the present invention can be administered by any known administration method.
  • the dosage of the compound pharmaceutical composition of the present invention can vary widely according to the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the administration route and the dosage form, etc.
  • a suitable daily dosage range of the compound of the present invention is 0.1-1000 mg/Kg body weight, preferably 1-500 mg/Kg body weight.
  • the above dosage may be administered in one dosage unit or divided into several dosage units, depending on the doctor's clinical experience and the administration regimen including the use of other treatments.
  • the compound or composition of the present invention can be taken alone or in combination with other therapeutic drugs or symptomatic drugs.
  • the compound of the present invention has a synergistic effect with other therapeutic drugs, its dosage should be adjusted according to the actual situation.
  • the fourth aspect of the technical solution of the present invention provides the use of the compound according to the first aspect of the present invention in the preparation of PARP-1/2 inhibitors, in the preparation of prevention and/or treatment of diseases related to PARP-1/2 Application in medicine, application in preparing anti-tumor medicine, application in preparing medicine for tumor-related diseases, wherein the tumor is selected from melanoma, gastric cancer, lung cancer, breast cancer, kidney cancer, liver cancer, Oral epidermal cancer, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, colon cancer, bladder cancer, glioma.
  • the compounds of this patent application exhibit very strong antitumor activity in vivo at low doses; and the compounds of this patent application have higher oral bioavailability than unsalted compounds.
  • the structure of the compound is determined by nuclear magnetic resonance (NMR) or high resolution mass spectrometry (HRMS).
  • NMR nuclear magnetic resonance
  • HRMS high resolution mass spectrometry
  • the NMR measurement is Varian mercury 300 or Varian mercury 400
  • the determination solvent is CDCl 3 , DMSO-d 6 , acetone-d 6 , CD 3 OD
  • the internal standard is TMS
  • the chemical shift is given in ppm.
  • mp is the melting point given in °C. The temperature is not corrected.
  • Silica gel column chromatography generally uses 200-300 mesh silica gel as a carrier.
  • NBS N-bromosuccinimide
  • DMAP 4-dimethylaminopyridine
  • AIBN azobisisobutyronitrile
  • Boc tert-butoxycarbonyl
  • HMDS Lithium hexamethyldisilazide
  • HBTU O-benzotriazole-tetramethylurea hexafluorophosphate
  • HATU 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate
  • HOBt 1-hydroxybenzotriazole
  • TFA trifluoroacetic acid
  • CD 3 OD deuterated methanol
  • TMS tetramethyl silicon
  • Collect human breast cancer MDA-MB-436 tumor cells under sterile conditions adjust the cell density to 1 ⁇ 10 7 cells/ml with sterile saline, take 0.2 ml and inoculate subcutaneously under the armpit and back of nude mice, wait for the tumor to grow to 1 cm The size was taken out under sterile conditions, cut into 1mm ⁇ 1mm tumor masses, and evenly inoculated under the skin of the axillary back of nude mice. After 7 days, when the tumor grows to 100-300 mm 3 , the animals are randomly divided into groups and started to be administered (recorded as day 0). Olaparib and the compound of Example 1 were administered orally daily.
  • the body weight was weighed twice a week and the length and width of the tumor were measured with a vernier caliper. After 23 days of administration, the nude mice were dislocated and sacrificed, and the tumor tissue was peeled off and weighed. Finally, calculate the tumor suppression rate, and evaluate the anti-tumor effect intensity with the tumor suppression rate.
  • Blank control group positive drug olaparib group (100mg/kg), Example 1 compound (1.1mg/kg) group, Example 1 compound (3.3mg/kg) group and Example 1 compound (10mg/kg) group.
  • Olaparib was suspended in a 0.5% sodium carboxymethyl cellulose solution, and the compound of Example 1 was dissolved in sterile water.
  • Tumor volume (TV): V 1/2 ⁇ a ⁇ b 2 , a and b represent the length and width of the tumor, respectively.
  • Inhibition rate (1-T/C) ⁇ 100, T is the weight of TV or tumor in the treatment group, and C is the weight of TV or tumor in the negative control group.
  • the compound of Example 1 has significant inhibitory activity on human breast cancer MDA-MB-436. The results are shown in Table 1.
  • Collect human breast cancer MDA-MB-436 tumor cells under sterile conditions adjust the cell density to 1 ⁇ 10 7 cells/ml with sterile saline, take 0.2 ml and inoculate subcutaneously under the armpit and back of nude mice, wait for the tumor to grow to 1 cm in diameter The size was taken out under sterile conditions, cut into tumors of 1mm ⁇ 1mm size, and evenly inoculated under the skin of the axillary back of nude mice. After 7 days, after the tumor grows to 100-300 mm 3 , the animals are randomly divided into groups and started to be administered (recorded as day 0). Olaparib and Compound P are administered orally daily.
  • Blank control group positive drug olaparib group (100mg/kg), compound P (25mg/kg) group, compound P (50mg/kg) group and compound P (100mg/kg) group.
  • Tumor volume (TV): V 1/2 ⁇ a ⁇ b 2 , a and b represent the length and width of the tumor, respectively.
  • Inhibition rate (1-T/C) ⁇ 100, T is the weight of TV or tumor in the treatment group, and C is the weight of TV or tumor in the negative control group.
  • Compound P can inhibit the growth of MDA-MB-436 tumor in a dose-dependent manner.
  • Compound P inhibits the growth of human breast cancer MDA-MB-436 in nude mice subcutaneous xenograft tumors
  • Example 1 Prototype compound P mother liquor (2.5 mg/mL) was diluted with acetonitrile containing internal standard propranolol to a working solution with a concentration of 5, 10, 25, 50, 100, 250, 500, 1000 ng/mL.
  • the prototype compound P of Example 1 and the compound of Example 1 were formulated with 0.5% CMC (containing Tween) as a 2.5 mg/mL suspension for oral administration, and the compound of Example 1 was formulated with 10% DMSO saline as a 0.25 mg/mL solution. Intravenously.
  • mice There were 26 mice, 5 in each group of oral group and 3 in each group of intravenous group. They were fasted for 12 hours before administration, and were free to drink water. Mice were bled from the orbital venous plexus at 5, 15, 30 min, 1, 2, 4, 6, 8, 12 h after oral administration of the prototype compound of Example 1 and the compound of Example 1 (25 mg/kg). Mice were bled from the orbital venous plexus 2, 5, 15, 30 min, 1, 2, 4, 6, 8, 12 h after intravenous injection of the compound of Example 1 (2.5 mg/10 mL/kg).
  • Mobile phase acetonitrile/water (containing 0.1% formic acid) gradient; flow rate: 0.2mL/min; MRM positive ion detection m/z 467.2 ⁇ 304.9 (YHP1318), m/z 260 ⁇ 183 (internal standard propranolol) .
  • the plasma pharmacokinetic parameters were calculated using WinNonlin software.
  • the peak area ratio of the test compound to the internal standard is taken as the ordinate, and the concentration of the test compound is taken as the abscissa. Correlation analysis is performed.
  • the prototype compound P (5-1000ng/ mL) The correlation between the concentration and the peak area ratio of the chromatogram is good, with a correlation coefficient >0.99.

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Abstract

本发明涉及含有哌嗪酮的喹唑啉-2,4-二酮盐类化合物作为PARP1/2抑制剂及其制备方法，含有所述化合物的药物组合物，以及所述化合物作为药物，尤其是作为抗肿瘤药物或作为肿瘤药物增敏剂与抗肿瘤药物联合应用。

Description

含有哌嗪酮的喹唑啉二酮盐类化合物、其制备方法、药物组合物和用途 技术领域
本发明涉及含有哌嗪酮的喹唑啉-2,4-二酮盐类化合物作为PARP1/2抑制剂及其制备方法，含有所述化合物的药物组合物，以及所述化合物作为药物，尤其是作为抗肿瘤药物或作为肿瘤药物增敏剂与抗肿瘤药物联合应用。
背景技术
多聚二磷酸腺苷核糖聚合酶(Poly ADP Ribose Polymerase，PARP)广泛存在于真核细胞中，是能够催化聚ADP核糖化的细胞核酶。该酶由Chambon等人于1963年首次报道，它具有保持染色体结构完整性、参与DNA的复制和转录、维持基因稳定性等重要作用[D’Amours D.Et al.Biochemical Journal,1999,342,249–268]。PARP家族中至少含有17个亚型，其中对PARP-1的研究最为广泛。[Ame J.C.et al.Bioessays,2004,26,882–893]。由于大多数化疗药物通过直接或间接地损伤DNA链起到杀伤肿瘤细胞的效果，抑制PARP-1的活性能有效阻滞损伤DNA的修复，从而与化疗药物产生协同抗肿瘤作用。PARP-1抑制剂对存在乳腺癌易感基因(BRCA-1、BRCA-2)突变细胞更为敏感，可单独用于BRCA突变肿瘤的治疗[Bryant,H.E.et al.Nature 2005,434,913-917].
在PARP家族中，PARP-2和PARP-1的同源性最高[Kutuzov,M.M.et al.Molecular Biology 48,485-495]。在生物学功能上，它和PARP-1既有相似之处，又有不同。和PARP-1一样，PARP-2也可以被损伤的DNA激活[Ame,J.C.et al.J Biol Chem 1999,274,17860-8]。通过对PARP-2基因敲除模型的观察，人们发现小鼠对电离辐射、甲基化试剂的敏感性升高，基因组的不稳定性增加。另外，PARP-2还具有独特的生理功能。对PARP-2基因进行下调，会阻止心肌细胞肥大[Geng,B.et al.Biochemical and Biophysical Research Communications 430,944-950]。在结肠炎小鼠模型中，对PARP-2基因进行敲除会改善炎症状况，并促进结肠功能的恢复[Popoff,I.et.al.Journal of Pharmacology and Experimental  Therapeutics 2002,303,1145-1154]。
目前已知的大多数PARP-1抑制剂对PARP-2也具有抑制作用，且活性相当。目前为止，已有奥拉帕尼、瑞卡帕尼和尼拉帕尼这三个PARP-1/2抑制剂分别于2014年、2016年和2017年作为抗肿瘤药物上市[Nature Reviews Drug Discovery,2017,16,229；Lin K.Y.et al.Cell,2017,169,183；Nature Biotechnology,2017,35,398]。
本专利设计合成了作用于PARP-1/2靶点的含有哌嗪酮的喹唑啉二酮盐类化合物，这类化合物相较于未成盐的原型化合物具有更好的体内抗肿瘤活性和口服生物利用度，旨在为治疗与PARP-1/2相关的疾病提供新的更加优良的物质基础。
发明内容
本发明解决的技术问题在于提供式I所示的含有哌嗪酮的喹唑啉-2,4(1H,3H)-二酮盐类衍生物、其制备方法、药物组合物、及其在制备PARP-1/2抑制剂及其潜在的药物中的用途、在制备抗肿瘤药物或抗肿瘤药物增敏剂中的用途。
为解决本发明的技术问题，本发明提供了如下技术方案：
本发明技术方案的第一方面是提供了如通式I所示的含有哌嗪酮的喹唑啉二酮盐类衍生物：
Figure PCTCN2019123295-appb-000001
在式I中，
R 1、R 2、R 3、R 4和R 5独立选自H、F、Cl、Br；
R 6选自如下原子或基团或结构片段：
(1)氢、取代或非取代的C1-8直链或支链烷基、取代或非取代的C2-8直链或支链烯基、取代或非取代的C2-8直链或支链炔基，其中取代基选自F、Cl、Br、 CN、ORc 1、SRc 2、NRc 3Rd 1、环丙基、环丙亚甲基、环丁基、氧杂环丁基、环戊基、环己基、Ar，其中所述的Rc 1、Rc 2、Rc 3、Rd 1独立地选自H、C1-4直链或支链烷基、环丙基、环丙亚甲基、环丁基，其中所述的Ar独立地选自取代或非取代的苯基、取代或非取代的含氮六元芳杂环、取代或非取代的五元芳杂环，其中取代基选自C1-4直链或支链烷基、卤素取代的C1-4直链或支链烷基、F、Cl、Br、NO 2、CN、亚甲二氧基、ORa′ 1、SRa′ 2、NRa′ 3Rb′ 1，其中所述的Ra′ 1、Ra′ 2、Ra′ 3、Rb′ 1独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；所述的苯环、含氮六元芳杂环、五元芳杂环上可以是单取代，也可以是多取代；六元芳杂环可以含有1个N原子，也可以含有多个氮原子；五元芳杂环可以含有一个杂原子，也可以含有多个杂原子，杂原子选自O,N,S；n选自1,2,3；其中所述的卤素包括F、Cl、Br；
(2)取代或非取代的C3-7环烷基、取代或非取代的3-8元环的氧杂环烷基、取代或非取代的3-8元环的氮杂环烷基，其中所述的取代基选自甲基、乙基、丙基、异丙基、F、Cl、Br、CN、ORc 1、SRc 2、NRc 3Rd 1、环丙基、环丙亚甲基、环丁基，其中所述的Rc 1、Rc 2、Rc 3、Rd 1、独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；3-8元环的氧杂环烷基和氮杂环烷基中可以含有1个杂原子，也可以同时含有多个杂原子；
(3)取代或非取代的苯基、取代或非取代的含氮六元芳杂环、取代或非取代的五元芳杂环，其中取代基选自C1-4直链或支链烷基、卤素取代的C1-4直链或支链烷基、F、Cl、Br、NO 2、CN、亚甲二氧基、ORa′ 1、SRa′ 2、NRa′ 3Rb′ 1，其中所述的Ra′ 1、Ra′ 2、Ra′ 3、Rb′ 1、独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；所述的苯环、含氮六元芳杂环、五元芳杂环上可以是单取代，也可以是多取代；六元芳杂环可以含有1个N原子，也可以含有多个氮原子；五元芳杂环可以含有一个杂原子，也可以含有多个杂原子，杂原子选自O,N,S；n选自1,2,3；其中所述的卤素包括F、Cl、Br；
M独立地选自不同碱金属(锂，钠，钾，铯)或是碱土金属盐(钙，镁，钡)。
在通式I中，所述的R 5独立地优选自F。
根据本发明通式I，本发明优选的化合物，包括但不限于通式I-A所示的化合物：
Figure PCTCN2019123295-appb-000002
在式I-A中，
R 1、R 2、R 3和R 4独立选自H、F、Cl、Br；
R′ 6选自如下原子或基团或结构片段：
(1)氢、取代或非取代的C1-8直链或支链烷基、取代或非取代的C2-8直链或支链烯基、取代或非取代的C2-8直链或支链炔基，其中取代基选自F、Cl、Br、CN、ORc 1、SRc 2、NRc 3Rd 1、环丙基、环丙亚甲基、环丁基、氧杂环丁基、环戊基、环己基、Ar，其中所述的Rc 1、Rc 2、Rc 3、Rd 1独立地选自H、C1-4直链或支链烷基、环丙基、环丙亚甲基、环丁基，其中所述的Ar独立地选自取代或非取代的苯基、取代或非取代的含氮六元芳杂环、取代或非取代的五元芳杂环，其中取代基选自C1-4直链或支链烷基、卤素取代的C1-4直链或支链烷基、F、Cl、Br、NO 2、CN、亚甲二氧基、ORa′ 1、SRa′ 2、NRa′ 3Rb′ 1，其中所述的Ra′ 1、Ra′ 2、Ra′ 3、Rb′ 1独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；所述的苯环、含氮六元芳杂环、五元芳杂环上可以是单取代，也可以是多取代；六元芳杂环可以含有1个N原子，也可以含有多个氮原子；五元芳杂环可以含有一个杂原子，也可以含有多个杂原子，杂原子选自O,N,S；n选自1,2,3；其中所述的卤素包括F、Cl、Br；
(2)取代或非取代的C3-7环烷基、取代或非取代的3-8元环的氧杂环烷基、取代或非取代的3-8元环的氮杂环烷基，其中所述的取代基选自甲基、乙基、丙基、异丙基、F、Cl、Br、CN、ORc 1、SRc 2、NRc 3Rd 1、环丙基、环丙亚甲基、环丁基，其中所述的Rc 1、Rc 2、Rc 3、Rd 1、独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；3-8元环的氧杂环烷基和氮杂环烷基中可以含有1个杂原子，也可以同时含有多个杂原子；
M独立地选自不同碱金属(锂，钠，钾，铯)或是碱土金属盐(钙，镁，钡)。
在通式I-A中，所述的R 1、R 2、R 3、R 4独立地优选自H、F。
根据本发明通式I，本发明优选的化合物，包括但不限于通式I-B所示的化合物：
Figure PCTCN2019123295-appb-000003
在式I-B中，
R 1、R 2、R 3、R 4独立选自H、F、Cl、Br；
Ar选自如下原子或基团或结构片段，包括
取代或非取代的苯基、取代或非取代的含氮六元芳杂环、取代或非取代的五元芳杂环，其中取代基选自C1-4直链或支链烷基、卤素取代的C1-4直链或支链烷基、F、Cl、Br、NO 2、CN、亚甲二氧基、ORa′ 1、SRa′ 2、NRa′ 3Rb′ 1，其中所述的Ra′ 1、Ra′ 2、Ra′ 3、Rb′ 1、独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；所述的苯环、含氮六元芳杂环、五元芳杂环上可以是单取代，也可以是多取代；六元芳杂环可以含有1个N原子，也可以含有多个氮原子；五元芳杂环可以含有一个杂原子，也可以含有多个杂原子，杂原子选自O,N,S；n选自1,2,3；其中所述的卤素包括F、Cl、Br；
M独立地选自不同碱金属(锂，钠，钾，铯)或是碱土金属盐(钙，镁，钡)。
在通式I-B中，所述的R 1、R 2、R 3、R 4、R 5独立地优选自H、F。
为完成本发明的目的，优选的化合物包括但不限定于：
1-(3-(4-戊烷-3’-基-3-氧代哌嗪)-1-甲酰基-4-氟-苄基)喹唑啉-2,4(1H,3H)-二酮钠盐
本发明技术方案的第二方面是提供了第一方面所述化合物的制备方法，采用 的技术方案包括以下步骤：R 1-R 4取代的不同喹唑啉二酮与包含R 5取代的3-甲氧羰基溴苄在HMDS的作用下分别发生选择性的烷基化反应，经水解后，包含R 1-R 5取代的不同的3-((2,4-二氧代-3,4-二氢喹唑啉-1(2H)-基)甲基)苯甲酸再与包含R 6的哌啶酮衍生物进行缩合，得到1-苄基喹唑啉二酮类化合物，最后与不同的碱发生酸碱反应得到1-苄基喹唑啉二酮类化合物的碱加成盐。其中所述的R 1、R 2、R 3、R 4、R 5、R 6、M的定义与本发明第一方面所述化合物的定义相同。
Figure PCTCN2019123295-appb-000004
试剂及反应条件:(a)脲,140℃,6h；(b)i)六甲基二硅基氨基锂(HMDS),浓硫酸,甲苯,回流,2h,ii)取代的5-(溴甲基)-2-氟苯甲酸甲酯,145℃,3h,iii)甲醇,己烷,70℃,30min；(c)LiOH,MeOH,H 2O,THF,55℃,2h；(d)2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU),1-羟基苯并三唑(HOBT),二异丙基乙胺(DIEA)，DMF(N,N-二甲基甲酰胺),overnight(过夜)；(e)MOH/H 2O/40℃；
其中所述的R 1、R 2、R 3、R 4、R 5、R 6和M的定义与本发明第一方面所述化合物的定义相同。
另外，上述反应中的起始原料及中间体容易得到，各步反应可依据已报道的文献或对本领域熟练技术人员来说可以用有机合成中的常规方法很容易合成。通式I所述化合物可以溶剂化物或非溶剂化物的形式存在，利用不同的溶剂进行结晶可能得到不同的溶剂化物。
本发明技术方案的第三方面是提供了一种药物组合物，所述药物组合物包括作为本发明技术方案第一方面所述的化合物和药学上的常用载体。
该组合物包括本发明中至少一种化合物和在药学上可接受的载体。所述的药物组合物选自片剂、胶囊、丸剂、注射剂、缓释制剂、控释制剂或各种微粒给药系统。该药物组合物可根据本领域公知的方法制备。可通过将本发明化合物与一种或多种药学上可接受的固体或液体赋形剂和/或辅剂结合，制成适于人或动物使用的任何剂型。本发明化合物在其药物组合物中的含量通常为0.1-95重量％。
本发明化合物或含有它的药物组合物可以单位剂量形式给药，给药途径可为肠道或非肠道，如口服、静脉注射、肌肉注射、皮下注射、鼻腔、口腔粘膜、眼、肺和呼吸道、皮肤、阴道、直肠等。
给药剂型可以是液体剂型、固体剂型或半固体剂型。液体剂型可以是溶液剂(包括真溶液和胶体溶液)、乳剂(包括o/w型、w/o型和复乳)、混悬剂、注射剂(包括水针剂、粉针剂和输液)、滴眼剂、滴鼻剂、洗剂和搽剂等；固体剂型可以是片剂(包括普通片、肠溶片、含片、分散片、咀嚼片、泡腾片、口腔崩解片)、胶囊剂(包括硬胶囊、软胶囊、肠溶胶囊)、颗粒剂、散剂、微丸、滴丸、栓剂、膜剂、贴片、气(粉)雾剂、喷雾剂等；半固体剂型可以是软膏剂、凝胶剂、糊剂等。
本发明化合物可以制成普通制剂、也制成是缓释制剂、控释制剂、靶向制剂及各种微粒给药系统。
这些制剂是按照本领域的技术人员所熟知的方法制备的。为制造片剂、胶囊剂、包衣剂所用的辅料是常规用的助剂，例如淀粉，明胶，阿拉伯胶，硅石，聚乙二醇，液体剂型所用的溶剂例如有水，乙醇，丙二醇，植物油类如玉米油，花生油，橄榄油等。含有本发明化合物的制剂中还可有其他助剂，例如表面活性剂，润滑剂，崩解剂，防腐剂，矫味剂，色素等。
为了将本发明化合物制成片剂，可以广泛使用本领域公知的各种赋形剂，包括稀释剂、黏合剂、润湿剂、崩解剂、润滑剂、助流剂。稀释剂可以是淀粉、糊精、蔗糖、葡萄糖、乳糖、甘露醇、山梨醇、木糖醇、微晶纤维素、硫酸钙、磷酸氢钙、碳酸钙等；湿润剂可以是水、乙醇、异丙醇等；粘合剂可以是淀粉浆、 糊精、糖浆、蜂蜜、葡萄糖溶液、微晶纤维素、阿拉伯胶浆、明胶浆、羧甲基纤维素钠、甲基纤维素、羟丙基甲基纤维素、乙基纤维素、丙烯酸树脂、卡波姆、聚乙烯吡咯烷酮、聚乙二醇等；崩解剂可以是干淀粉、微晶纤维素、低取代羟丙基纤维素、交联聚乙烯吡咯烷酮、交联羧甲基纤维素钠、羧甲基淀粉钠、碳酸氢钠与枸橼酸、聚氧乙烯山梨糖醇脂肪酸酯、十二烷基磺酸钠等；润滑剂和助流剂可以是滑石粉、二氧化硅、硬脂酸盐、酒石酸、液体石蜡、聚乙二醇等。
还可以将片剂进一步制成包衣片，例如糖包衣片、薄膜包衣片、肠溶包衣片，或双层片和多层片。
为了将给药单元制成胶囊剂，可以将有效成分本发明化合物与稀释剂、助流剂混合，将混合物直接置于硬胶囊或软胶囊中。也可将有效成分本发明化合物先与稀释剂、黏合剂、崩解剂制成颗粒或微丸，再置于硬胶囊或软胶囊中。用于制备本发明化合物片剂的各稀释剂、黏合剂、润湿剂、崩解剂、助流剂品种也可用于制备本发明化合物的胶囊剂。
为将本发明化合物制成注射剂，可以用水、乙醇、异丙醇、丙二醇或它们的混合物作溶剂并加入适量本领域常用的增溶剂、助溶剂、pH调剂剂、渗透压调节剂。增溶剂或助溶剂可以是泊洛沙姆、卵磷脂、羟丙基-β-环糊精等；pH调剂剂可以是磷酸盐、醋酸盐、盐酸、氢氧化钠等；渗透压调节剂可以是氯化钠、甘露醇、葡萄糖、磷酸盐、醋酸盐等。如制备冻干粉针剂，还可加入甘露醇、葡萄糖等作为支撑剂。
此外，如需要，也可以向药物制剂中添加着色剂、防腐剂、香料、矫味剂或其它添加剂。
为达到用药目的，增强治疗效果，本发明的药物或药物组合物可用任何公知的给药方法给药。
本发明化合物药物组合物的给药剂量依照所要预防或治疗疾病的性质和严重程度，患者或动物的个体情况，给药途径和剂型等可以有大范围的变化。一般来讲，本发明化合物的每天的合适剂量范围为0.1-1000mg/Kg体重，优选为1-500mg/Kg体重。上述剂量可以一个剂量单位或分成几个剂量单位给药，这取决于医生的临床经验以及包括运用其它治疗手段的给药方案。
本发明的化合物或组合物可单独服用，或与其他治疗药物或对症药物合并 使用。当本发明的化合物与其它治疗药物存在协同作用时，应根据实际情况调整它的剂量。
本发明技术方案的第四方面是提供了本发明第一方面所述的化合物在制备PARP-1/2抑制剂中的应用、在制备预防和\或治疗与PARP-1/2有关的疾病的药物中的应用、在制备抗肿瘤药物中的应用、在制备与肿瘤有关的疾病的药物中的应用，其中，所述的肿瘤选自黑色素瘤、胃癌、肺癌、乳腺癌、肾癌、肝癌、口腔表皮癌、宫颈癌、卵巢癌、胰腺癌、前列腺癌、结肠癌、膀胱癌、神经胶质瘤。
有益技术效果：
本专利申请化合物在低剂量时，呈现非常强的体内抗肿瘤活性；且本专利申请的化合物相较于未成盐化合物具有更高的口服生物利用度。
具体实施方式
以下将结合实施例对发明做进一步说明，但并不限制本发明的范围。
化合物的结构是通过核磁共振(NMR)或高分辨质谱(HRMS)来确定的。NMR的测定是用Varian mercury 300或者Varian mercury 400，测定溶剂为CDCl 3、DMSO-d 6、acetone-d 6、CD 3OD，内标为TMS，化学位移是以ppm作为单位给出。m.p.是以℃给出的熔点，温度未加校正。硅胶柱层析一般使用200～300目硅胶为载体。
缩写列表：
TLC:薄层色谱；
CDCl 3：氘代氯仿；DIEA：二异丙基乙胺；TFA：三氟乙酸；TEA：三乙胺
DMF：N,N-二甲基甲酰胺；THF：四氢呋喃；PE：石油醚；EA：乙酸乙酯
min：分钟；r.t.室温；DCM：二氯甲烷；h：小时；Et 3N：三乙胺
EDC或EDCI：1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐
NBS：N-溴代丁二酰亚胺；DMAP：4-二甲氨基吡啶
AIBN：偶氮二异丁腈；Boc：特丁氧羰基
HMDS：六甲基二硅基氨基锂
HBTU：O-苯并三氮唑-四甲基脲六氟磷酸酯
HATU：2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯
HOBt：1-羟基苯并三氮唑；TFA：三氟乙酸；
Et 3N：三乙胺；HOBt：1-羟基苯并三唑；NaH：氢化钠；
CCl 4：四氯化碳；DMSO-d 6：氘代二甲基亚砜；acetone-d 6：氘代丙酮
CD 3OD：氘代甲醇；TMS：四甲基硅
中间体的制备：
(一)5-((2,4-二氧代-3,4-二氢喹唑啉-1(2H)-基)甲基)-2-氟苯甲酸的制备
Figure PCTCN2019123295-appb-000005
a)2-氟-5-溴甲基苯甲酸甲酯
将2-氟-5-甲基苯甲酸甲酯(6.72g,40mmol)加入反应瓶中，加入CCl 4(100mL)，NBS(7.47g,42mmol)，AIBN(1.29g,8mmol)，乙腈(20mL)，加热回流反应，1.5h后停止反应，加入DCM(100mL)，用饱和NaCl(50mL×3)洗，无水硫酸镁干燥，柱层析(EA:P＝1:150)，得到白色固体6.4g，产率65％。
1H NMR(300MHz,CDCl 3)δ(ppm):7.96-7.99(1H,m),7.53-7.58(1H,m),7.10-7.17(1H,m),4.48(2H,s),3.94(3H,s).
b)5-((2,4-二氧代-3,4-二氢喹唑啉-1(2H)-基)甲基)-2-氟苯甲酸甲酯
将喹唑啉-2,4(1H,3H)-二酮(2g,12.3mmol)加入反应瓶中，加入甲苯(40mL)，加入HMDS(10.4mL,61.6mmol)，浓硫酸(20滴)，加热回流反应，5h后反应液澄清，浓缩，向反应残渣中加入2-氟-5-溴甲基苯甲酸甲酯，加热至140℃反应，4h后停止反应，降温至100℃，加入二氧六环(15mL)，甲醇(10mL)，70℃下搅拌30min，降温至0℃,抽滤，得到白色固体3.3g，产率81.8％。
1H NMR(300MHz,DMSO-d 6)δ(ppm):11.47(brs,1H),8.02(d,J＝5.7Hz,1H),7.86(d,J＝4.5Hz,1H),7.66(t,J＝5.7Hz,1H),7.58-7.61(m,1H),7.23-7.34(m,3H),5.34(s,2H),3.83(s,3H)；m.p.212-214℃.
c)将5-((2,4-二氧代-3,4-二氢喹唑啉-1(2H)-基)甲基)-2-氟苯甲酸甲酯(2g,6.1mmol)加入甲醇(30mL)，THF(15mL)，将(421mg,18.3mmol)氢氧化锂溶于(15mL)水中加入反应瓶中，升温至50℃反应，30min后停止反应，浓缩，水层用稀HCl溶液调pH值至2左右，抽滤，滤饼水洗，得到白色固体1.4g，产率73.2％。
1H NMR(300MHz,DMSO-d 6)δ(ppm):13.31(s,1H),11.76(s,1H),8.02(d,J＝5.7Hz,1H),7.82(d,J＝4.2Hz,1H),7.66(t,J＝5.7Hz,1H),7.55-7.57(m,1H),7.23-7.31(m,3H),5.33(s,2H).m.p.>250℃
(二)N-苄氧羰酰基-3-氧代哌嗪的制备
Figure PCTCN2019123295-appb-000006
将哌嗪2-酮(1g,10mmol)，加入乙酸乙酯(40mL)，水(20mL)中，室温搅拌下加入K 2CO 3(6.9g,50mmol)，将苄氧羰酰氯(2.1mL,15mmol)滴加入反应瓶中，滴毕，室温搅拌反应，次日停止反应，分层，有机层用饱和NaCl溶液20mL×2洗，无水硫酸镁干燥，柱层析(D:M＝75:1)得到白色固体1.4g，产率59.8％。
1H-NMR(400MHz,CDCl 3)δ(ppm):7.32-7.40(m,5H),6.94(brs,1H),5.16(s,2H),4.16(s,2H),3.70(t,J＝4.8Hz,2H),3.40(brs,2H)
实施例1
1-(3-(4-戊烷-3’-基-3-氧代哌嗪)-1-甲酰基-4-氟-苄基)喹唑啉-2,4(1H,3H)-二酮钠盐
Figure PCTCN2019123295-appb-000007
a)苄基4-戊基3’-基-3-氧代哌嗪-1-甲酸酯
将N-苄氧羰酰基-3-氧代哌嗪(8g,10.25mmol)加入DMF(80mL)，氩气保护下， 室温下加入NaH(2.7g,68.33mmol)，室温搅拌1h，加入3-溴戊烷(10.56mL,84.29mmol)，升温至50℃反应，3d后停止反应，加入水，用乙酸乙酯200mL萃取，有机层用饱和NaCl溶液(50mL×2)洗，无水硫酸镁干燥，柱层析，得到产物4.6g，产率44.3％。
1H-NMR(400MHz,CDCl 3)δ(ppm):7.28-7.36(m,5H),5.16(s,1H),4.45-4.50(m,1H),4.19(s,2H),3.68(t,J＝5.2Hz,2H),3.16(brs,2H),1.35-1.58(m,4H),0.85(t,J＝7.6Hz,6H),
b)1-戊烷-3’-基哌嗪-2-酮
将苄基4-戊基3’-基-3-氧代哌嗪-1-甲酸酯(4.5g)，加入EtOH(60mL)，加入10％Pd/C(1.35g)，常温常压下氢化反应，4h后停止反应，过滤，浓缩，得到无色油状物2.5g，产率99.6％。
c)1-(3-(4-戊烷-3’-基-3-氧代哌嗪)-1-甲酰基-4-氟-苄基)喹唑啉-2,4(1H,3H)-二酮(化合物P)
将5-((2,4-二氧代-3,4-二氢喹唑啉-1(2H)-基)甲基)-2-氟苯甲酸(3.14g,10mmol)，加入DMF(60mL)，EDC(3.84g,20mmol)，HOBt(2.7g,20mmol)，DIEA(5.23mL,30mmol)以及化合物1-戊烷-3’-基哌嗪-2-酮(2.5g,14.7mmol)，室温搅拌反应，次日停止反应，加水，用MeOH:DCM＝1:10的混合液萃取，有机层用水(50mL×2)洗，饱和NaCl溶液(50mL×2)洗，无水硫酸镁干燥，用乙酸乙酯，甲醇和正己烷重结晶得到产物3.25g，总产率69.7％。
1H-NMR(400MHz,CDCl 3)δ(ppm):10.80(s,0.4H),10.67(s,0.6H),8.46(d,J＝6.8Hz,1H),7.69-7.81(m,3H),7.32-7.39(m,2H),7.04(t,J＝9.2Hz,1H),4.42-4.51(m,2H),4.28(s,2H),4.04(s,1H),3.96(s,1H),3.52(s,1H),3.24(t,J＝5.2Hz,1H),3.11(s,1H),1.37-1.59(m,4H),0.82-0.86(m,6H).
d)1-(3-(4-戊烷-3’-基-3-氧代哌嗪)-1-甲酰基-4-氟-苄基)喹唑啉-2,4(1H,3H)-二酮钠盐(实施例1化合物)
将化合物1-(3-(4-戊烷-3’-基-3-氧代哌嗪)-1-甲酰基-4-氟-苄基)喹唑啉-2,4(1H,3H)-二酮(1g,2.14mmol)置于反应瓶中，NaOH(93.3mg,2.33mmol)溶于2mL水中，加入反应瓶中，升温至40℃反应1h，至反应液澄清，浓缩至干，用乙酸乙酯和少量甲醇溶解后过滤，滤液浓缩，用乙酸乙酯，甲醇和正己烷重结晶得到白色固体720mg，产率68.9％。
1H-NMR(400MHz,DMSO-d 6)δ(ppm):7.87-7.91(m,1H),7.35-7.45(m,2H),7.22-7.30(m,2H),6.97-7.03(m,2H),5.27(s,2H),4.17-4.27(m,2H),3.80-3.83(m,1H),3.19-3.21(m,1H),2.94-2.97(m,1H),2.49-2.51(m,2H),1.09-1.47(m,4H),0.74(t,J＝7.6Hz,6H).
药理实验：
实验例1：动物体内药效学实验：
(一)实施例1化合物对人乳腺癌MDA-MB-436皮下移植瘤的抑制作用
实验方法：
1.步骤
无菌条件下收集人乳腺癌MDA-MB-436肿瘤细胞，用灭菌生理盐水调整细胞密度至1×10 7个/ml，取0.2ml接种于裸鼠腋背部皮下，待肿瘤生长至直径1cm大小，无菌条件下取出，切成1mm×1mm大小的瘤块，均匀接种于裸鼠腋背部皮下。7日后待肿瘤生长至100～300mm 3后，将动物随机分组，开始给药(记为第0天)。奥拉帕尼和实施例1化合物每天口服给药。每周两次称量体重并用游标卡尺测量肿瘤的长度和宽度，给药23天后将裸鼠脱臼处死，剥离肿瘤组织，称重。最后计算肿瘤抑制率，以肿瘤抑制率评价抗肿瘤作用强度。
2.分组
空白对照组，阳性药奥拉帕尼组(100mg/kg)，实施例1化合物(1.1mg/kg)组，实施例1化合物(3.3mg/kg)组和实施例1化合物(10mg/kg)组。
3.药物配制
奥拉帕尼混悬于0.5％羧甲基纤维素钠溶液中，实施例1化合物溶于灭菌水中。
4.计算方法
肿瘤体积(TV)：V＝1/2×a×b 2，a和b分别表示瘤体长和宽。
抑瘤率：Inhibition(％)＝(1-T/C)×100，T为治疗组TV或肿瘤重量，C为阴性对照组TV或肿瘤重量。
实验结果：
实施例1化合物对人乳腺癌MDA-MB-436具有显著的抑制活性，结果见表1。
表1.实施例1化合物对人乳腺癌MDA-MB-436在裸鼠皮下异体移植瘤的生长抑制作用
Figure PCTCN2019123295-appb-000008
与对照组比较，**P<0.01
NA:不适用
(二)实施例1原型化合物P对人乳腺癌MDA-MB-436裸鼠皮下移植瘤的抑制作用
实验方法：
1.步骤
无菌条件下收集人乳腺癌MDA-MB-436肿瘤细胞，用灭菌生理盐水调整细胞密度至1×10 7个/ml，取0.2ml接种于裸鼠腋背部皮下，待肿瘤生长至直径1cm大小，无菌条件下取出，切成1mm×1mm大小的瘤块，均匀接种于裸鼠腋背部皮下。7日后待肿瘤生长至100～300mm 3后，将动物随机分组，开始给药(记为第0天)。奥拉帕尼和化合物P每天口服给药。每周两次称量体重并用游标卡尺测量肿瘤的长度和宽度，给药18天后将裸鼠脱臼处死，剥离肿瘤组织，称重。最后计算肿瘤抑制率，以肿瘤抑制率评价抗肿瘤作用强度。
2.分组
空白对照组，阳性药奥拉帕尼组(100mg/kg)，化合物P(25mg/kg)组，化合物P(50mg/kg)组和化合物P(100mg/kg)组。
3.药物配制
所有化合物均混悬于0.5％羧甲基纤维素钠溶液中。
4.计算方法
肿瘤体积(TV)：V＝1/2×a×b 2，a和b分别表示瘤体长和宽。
抑瘤率：Inhibition(％)＝(1-T/C)×100，T为治疗组TV或肿瘤重量，C为阴性对照组TV或肿瘤重量。
实验结果：化合物P可呈剂量依赖性抑制MDA-MB-436肿瘤的生长。
表2.化合物P对人乳腺癌MDA-MB-436在裸鼠皮下异体移植瘤的生长抑制作用
Figure PCTCN2019123295-appb-000009
与对照组比较，***P<0.001
NA:不适用
实验例2 药代实验
实验方法
1.血浆样品标准曲线建立
实施例1原型化合物P母液(2.5mg/mL)用含内标普萘洛尔乙腈稀释成浓 度为5、10、25、50、100、250、500、1000ng/mL工作液。
空白小鼠血浆30μL分别加入不同浓度实施例1原型化合物工作液30μL和含内标(普萘洛尔，0.2μg/mL)乙腈30μL，混旋后离心(14000rpm×5min)两次，取上清液5μL进行LC/MS/MS分析。
2.小鼠口服/静脉注射实施例1原型化合物P和实施例1化合物的血浆药代动力学研究
实施例1原型化合物P和实施例1化合物以0.5％CMC(含吐温)配制成2.5mg/mL悬液用于口服，实施例1化合物以10％DMSO生理盐水配制成0.25mg/mL溶液用于静脉注射。
小鼠26只，口服组每组5只，静脉组每组3只，采用分段采血法。给药前均禁食12h，自由饮水。小鼠口服实施例1原型化合物和实施例1化合物(25mg/kg)后5、15、30min、1、2、4、6、8、12h自眼眶静脉丛取血。小鼠静脉注射实施例1化合物(2.5mg/10mL/kg)后2、5、15、30min、1、2、4、6、8、12h自眼眶静脉丛取血。分离血浆30μL，加入含内标普萘洛尔的乙腈60μL，混旋后离心(14000rpm×5min)两次，取上清液5μL进行LC/MS/MS分析。超标线样品进行稀释后测定。
3.LC/MS/MS条件
色谱柱：Zobax C18(100mm×2.1mm，3.5μm)；柱温：37℃，
流动相：乙腈/水(含0.1％甲酸)梯度；流速：0.2mL/min；MRM正离子方式检测m/z 467.2→304.9(YHP1318)，m/z 260→183(内标普萘洛尔)。
4.数据分析
应用WinNonlin软件计算血浆药代动力学参数。
实验结果：
1.血浆样品标准曲线
根据各样品LC/MS/MS图谱，以待测化合物与内标的峰面积比为纵坐标，待测化合物浓度为横坐标，进行相关分析，血浆样品中实施例1原型化合物P(5-1000ng/mL)浓度与色谱峰面积比相关关系良好，相关系数>0.99。
2.小鼠口服/静脉注射实施例1原型化合物P和实施例1化合物的血浆药代动力学研究
小鼠口服及静脉注射实施例1原型化合物P和实施例1后，药代动力学参数 见表3。小鼠口服实施例1化合物的生物利用度高于其原型化合物P。
表3小鼠口服和静脉注射实施例1化合物及化合物P的血浆药代动力学参数
Figure PCTCN2019123295-appb-000010

Claims (11)

  1. 如通式I所示的含有哌嗪酮的喹唑啉二酮盐类化合物，
    Figure PCTCN2019123295-appb-100001
    在式I中，
    R 1、R 2、R 3、R 4和R 5独立选自H、F、Cl、Br；
    R 6选自如下原子或基团或结构片段：
    (1)氢、取代或非取代的C1-8直链或支链烷基、取代或非取代的C2-8直链或支链烯基、取代或非取代的C2-8直链或支链炔基，其中取代基选自F、Cl、Br、CN、ORc 1、SRc 2、NRc 3Rd 1、环丙基、环丙亚甲基、环丁基、氧杂环丁基、环戊基、环己基、Ar，其中所述的Rc 1、Rc 2、Rc 3、Rd 1独立地选自H、C1-4直链或支链烷基、环丙基、环丙亚甲基、环丁基，其中所述的Ar独立地选自取代或非取代的苯基、取代或非取代的含氮六元芳杂环、取代或非取代的五元芳杂环，其中取代基选自C1-4直链或支链烷基、卤素取代的C1-4直链或支链烷基、F、Cl、Br、NO 2、CN、亚甲二氧基、ORa′ 1、SRa′ 2、NRa′ 3Rb′ 1，其中所述的Ra′ 1、Ra′ 2、Ra′ 3、Rb′ 1独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；所述的苯环、含氮六元芳杂环、五元芳杂环上可以是单取代，也可以是多取代；六元芳杂环可以含有1个N原子，也可以含有多个氮原子；五元芳杂环可以含有一个杂原子，也可以含有多个杂原子，杂原子选自O,N,S；n选自1,2,3；其中所述的卤素包括F、Cl、Br；
    (2)取代或非取代的C3-7环烷基、取代或非取代的3-8元环的氧杂环烷基、取代或非取代的3-8元环的氮杂环烷基，其中所述的取代基选自甲基、乙基、丙基、异丙基、F、Cl、Br、CN、ORc 1、SRc 2、NRc 3Rd 1、环丙基、环丙亚甲基、环丁基，其中所述的Rc 1、Rc 2、Rc 3、Rd 1、独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；3-8元环的氧杂环烷基和氮杂环烷基中可以 含有1个杂原子，也可以同时含有多个杂原子；
    (3)取代或非取代的苯基、取代或非取代的含氮六元芳杂环、取代或非取代的五元芳杂环，其中取代基选自C1-4直链或支链烷基、卤素取代的C1-4直链或支链烷基、F、Cl、Br、NO 2、CN、亚甲二氧基、ORa′ 1、SRa′ 2、NRa′ 3Rb′ 1，其中所述的Ra′ 1、Ra′ 2、Ra′ 3、Rb′ 1、独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；所述的苯环、含氮六元芳杂环、五元芳杂环上可以是单取代，也可以是多取代；六元芳杂环可以含有1个N原子，也可以含有多个氮原子；五元芳杂环可以含有一个杂原子，也可以含有多个杂原子，杂原子选自O,N,S；n选自1,2,3；其中所述的卤素包括F、Cl、Br；
    M独立地选自碱金属或碱土金属。
  2. 根据权利要求1的含有哌嗪酮的喹唑啉二酮盐类化合物，其特征在于，所述的喹唑啉二酮盐类化合物如通式IA所示，
    Figure PCTCN2019123295-appb-100002
    在式IA中，
    R 1、R 2、R 3和R 4独立选自H、F、Cl、Br；
    R′ 6选自如下原子或基团或结构片段：
    (1)氢、取代或非取代的C1-8直链或支链烷基、取代或非取代的C2-8直链或支链烯基、取代或非取代的C2-8直链或支链炔基，其中取代基选自F、Cl、Br、CN、ORc 1、SRc 2、NRc 3Rd 1、环丙基、环丙亚甲基、环丁基、氧杂环丁基、环戊基、环己基、Ar，其中所述的Rc 1、Rc 2、Rc 3、Rd 1独立地选自H、C1-4直链或支链烷基、环丙基、环丙亚甲基、环丁基，其中所述的Ar独立地选自取代或非取代的苯基、取代或非取代的含氮六元芳杂环、取代或非取代的五元芳杂环，其中取代基选自C1-4直链或支链烷基、卤素取代的C1-4直链或支链烷基、F、 Cl、Br、NO 2、CN、亚甲二氧基、ORa′ 1、SRa′ 2、NRa′ 3Rb′ 1，其中所述的Ra′ 1、Ra′ 2、Ra′ 3、Rb′ 1独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；所述的苯环、含氮六元芳杂环、五元芳杂环上可以是单取代，也可以是多取代；六元芳杂环可以含有1个N原子，也可以含有多个氮原子；五元芳杂环可以含有一个杂原子，也可以含有多个杂原子，杂原子选自O,N,S；n选自1,2,3；其中所述的卤素包括F、Cl、Br；
    (2)取代或非取代的C3-7环烷基、取代或非取代的3-8元环的氧杂环烷基、取代或非取代的3-8元环的氮杂环烷基，其中所述的取代基选自甲基、乙基、丙基、异丙基、F、Cl、Br、CN、ORc 1、SRc 2、NRc 3Rd 1、环丙基、环丙亚甲基、环丁基，其中所述的Rc 1、Rc 2、Rc 3、Rd 1、独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚甲基、环丁基；3-8元环的氧杂环烷基和氮杂环烷基中可以含有1个杂原子，也可以同时含有多个杂原子；
    M独立地选自碱金属或碱土金属。
  3. 根据权利要求1的含有哌嗪酮的喹唑啉二酮盐类化合物，其特征在于，所述的喹唑啉二酮盐类化合物如通式IB所示
    Figure PCTCN2019123295-appb-100003
    在式IB中，
    R 1、R 2、R 3、R 4独立选自H、F、Cl、Br；
    Ar选自如下原子或基团或结构片段，包括
    取代或非取代的苯基、取代或非取代的含氮六元芳杂环、取代或非取代的五元芳杂环，其中取代基选自C1-4直链或支链烷基、卤素取代的C1-4直链或支链烷基、F、Cl、Br、NO 2、CN、亚甲二氧基、ORa′ 1、SRa′ 2、NRa′ 3Rb′ 1，其中所述的Ra′ 1、Ra′ 2、Ra′ 3、Rb′ 1、独立地选自H、甲基、乙基、丙基、异丙基、环丙基、环丙亚 甲基、环丁基；所述的苯环、含氮六元芳杂环、五元芳杂环上可以是单取代，也可以是多取代；六元芳杂环可以含有1个N原子，也可以含有多个氮原子；五元芳杂环可以含有一个杂原子，也可以含有多个杂原子，杂原子选自O,N,S；n选自1,2,3；其中所述的卤素包括F、Cl、Br；
    M独立地选自碱金属或是碱土金属。
  4. 根据权利要求1-3任一项的含有哌嗪酮的喹唑啉二酮盐类化合物，其特征在于，所述的M选自锂、钠、钾、铯、钙、镁、钡。
  5. 根据权利要求1的含有哌嗪酮的喹唑啉二酮盐类化合物，所述的喹唑啉二酮盐类化合物如下：
    1-(3-(4-戊烷-3’-基-3-氧代哌嗪)-1-甲酰基-4-氟-苄基)喹唑啉-2,4(1H,3H)-二酮钠盐。
  6. 权利要求1-5任一项含有哌嗪酮的喹唑啉二酮盐类化合物的制备方法，其特征在于，包括如下步骤：
    R 1-R 4取代的不同喹唑啉二酮与包含R 5取代的3-甲氧羰基溴苄在HMDS的作用下分别发生选择性的烷基化反应，经水解后，包含R 1-R 5取代的不同的3-((2,4-二氧代-3,4-二氢喹唑啉-1(2H)-基)甲基)苯甲酸再与包含R 6的哌啶酮衍生物进行缩合，得到1-苄基喹唑啉二酮类化合物，最后与不同的碱发生酸碱反应得到1-苄基喹唑啉二酮类化合物的碱加成盐；其中所述的R 1、R 2、R 3、R 4、R 5、R 6、M的定义与权利要求1-5任一项的定义相同，
    Figure PCTCN2019123295-appb-100004
    试剂及反应条件:(a)脲,140℃,6h；(b)i)六甲基二硅基氨基锂(HMDS),浓硫酸,甲苯,回流,2h,ii)取代的5-(溴甲基)-2-氟苯甲酸甲酯,145℃,3h,iii)甲醇,己烷,70℃,30min；(c)LiOH,MeOH,H 2O,THF,55℃,2h；(d)2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU),1-羟基苯并三唑(HOBT),二异丙基乙胺(DIEA)，DMF(N,N-二甲基甲酰胺),overnight(过夜)；(e)MOH/H 2O/40℃；
    其中所述的R 1、R 2、R 3、R 4、R 5、R 6、M的定义如权利要求1-5任一项所述。
  7. 一种药物组合物，其特征在于，包含有效剂量的权利要求1-5中任一项的含有哌嗪酮的喹唑啉二酮盐类化合物和药效学上可接受的载体。
  8. 权利要求1-5任一项的含有哌嗪酮的喹唑啉二酮盐类化合物在制备PARP-1/2抑制剂中的应用。
  9. 权利要求1-5中任一项的含有哌嗪酮的喹唑啉二酮盐类化合物在制备预防和\或治疗PARP-1/2有关的疾病的药物中的应用。
  10. 权利要求1-5中任一项的含有哌嗪酮的喹唑啉二酮盐类化合物在制备抗肿瘤 药物或抗肿瘤药物增敏剂中的应用。
  11. 根据权利要求10的应用，其特征在于，所述的肿瘤选自黑色素瘤、胃癌、肺癌、乳腺癌、三阴乳腺癌、肾癌、肝癌、口腔表皮癌、宫颈癌、卵巢癌、胰腺癌、前列腺癌、结肠癌、膀胱癌、神经胶质瘤。
PCT/CN2019/123295 2018-12-05 2019-12-05 含有哌嗪酮的喹唑啉二酮盐类化合物、其制备方法、药物组合物和用途 WO2020114455A1 (zh)

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