CN110627801A - HDAC inhibitor and application thereof - Google Patents

HDAC inhibitor and application thereof Download PDF

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CN110627801A
CN110627801A CN201910992789.6A CN201910992789A CN110627801A CN 110627801 A CN110627801 A CN 110627801A CN 201910992789 A CN201910992789 A CN 201910992789A CN 110627801 A CN110627801 A CN 110627801A
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hdac
inhibitor
dioxa
benzo
substituted
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CN110627801B (en
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马晓东
王昊
方方
陶强强
彭成军
孙松
王虎传
胡海霞
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Anhui University of Traditional Chinese Medicine AHUTCM
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HEFEI YUANZHI PHARMACEUTICAL R & D Co Ltd
Anhui University of Traditional Chinese Medicine AHUTCM
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Abstract

The invention discloses an HDAC inhibitor, which is a bifendate derivative shown in general formulas (I) - (IV), a pharmaceutically acceptable salt or a deuteron thereof, and a pharmaceutical composition containing the same, and application of the inhibitor in preparing anti-tumor, autoimmune disease, inflammation or Alzheimer disease drugs. The bifendate derivatives prepared by the invention have HDAC (histone deacetylase) inhibitory activity, and pharmacodynamic experiments show that the compounds can be used as medicaments for treating tumors, autoimmune diseases, inflammations or Alzheimer's disease.

Description

HDAC inhibitor and application thereof
Technical Field
The invention relates to the field of pharmaceutical chemistry, in particular to bifendate derivatives with HDAC (HDAC) inhibitory activity and a preparation method and application thereof.
Background
Histone Deacetylases (HDACs) play an important role in epigenetic regulation, and can regulate gene expression by catalyzing the deacetylation process of Histone or non-Histone. Of the four subcategories of HDACs, class I HDACs are closely related to tumorigenesis and development and are found in a wide variety of organs of the human body. The study demonstrated that HDAC1 subtype in HDACs class I is highly expressed in a variety of malignancies, such as endometrial, lung, prostate, breast, colon, Pancreatic, blood cancers, etc. (Zhu S, et al, Mir-548an, transcribed descending by HIF 1. alpha./HDAC 1, supra viral genetics of pathological Cancer by targeting expression. mol Cancer Ther,2016,15 (2209): 9-; Zhao R, et al, DNA damage-binding reactions HDAC1to expression Bcl-2transcription initiation man cell. mol Cancer, 2014,12(3): 370). High expression of HDAC1 can cause abnormal histone acetylation, and is closely related to tumor development and development, so the goal of treating tumor can be achieved by inhibiting HDAC1 (Stubbs, M.C, et al. selective Inhibition of HDAC1 and HDAC2 as aPotent Therapeutic Option for B-ALL. clin Cancer Res,2015,21(10): 2348-.
The HDAC6 subtype in class II HDACs has unique structural and substrate specificity, and its expression and functional changes are closely related to various diseases. The research shows that the abnormal expression of HDAC6 can promote the canceration of cells and the growth, proliferation and angiogenesis of tumor cells by regulating Ras, EGFR and other signal pathways; in addition, its high expression also enhances the metastatic and invasive properties of tumor cells (Lee YS, et al. the cytoplasmic deacylase HDAC6is required for clinical oncogenic clinical diagnosis. cancer Res,2008,68: 7561-. Meanwhile, HDAC6is also closely related to pathological processes of neurodegenerative diseases (such as Alzheimer disease), inflammation, autoimmune diseases, etc. (Selenica M L, et al, Histone deacetylase 6inhibition, memories and tau repair tools Resther,2014,6(1): 12; de Zoeten E F, et al, Histone deacetylase 6and Heat repair tools 90control the functions of Foxp3(+) T-regulation cells mol, 2011,31(10): 2066-2078; Rega N L et al, specificity HDAC 6inhibition, Y-transcription of acids and polypeptides, SLE W2016,162: N L, et al, specificity HDAC 6inhibition, SLE-transcription kinase, SLE 73, SLE 3, S5, S.
HDACs inhibitors which are now marketed mainly are broad-spectrum HDACs inhibitors, including vorinostat (SAHA), belinostat and panobinostat, but they have side effects such as nausea, vomiting, myelosuppression and prolongation of the QT interval, and their range of application is limited, so there is a strong need to find new HDAC inhibitors to improve the safety of this class of drugs (Balasuramanian S, et al, Isooform-specific histone deacetylase inhibitors: the next step? Cancer Lett,2009,280:211 @, 221.). subtype-selective HDAC inhibitors are currently the focus of research, in which HDAC6 selective inhibitors are of interest, but no HDAC drugs are marketed, only ACY-1215 are in clinical studies, besides direct intervention in the life activities of tumor cells, HDAC6 selective inhibitors may also down-regulate Programmed broad-spectrum Death ligands (Programmed Death-platelet-PD-1, thus exerting more selective action on HDAC inhibitors than HDAC inhibitors such as HDAC6, CD 6, HDAC inhibitors found to produce more selective effects than HDAC inhibitors for clinical therapy such as HDAC6, 16, 6, and multiple HDAC inhibitors.
Bifendate is a structural analogue of Chinese medicine monomer schisandrin and is clinically used as a liver protection medicine. In recent years, research and research have been conducted on the application of the compounds in the anti-tumor field. However, to date, no HDAC inhibitors based on the biphenyldicarboxylate parent nucleus have been reported.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a class of HDAC inhibitors and application thereof.
The technical scheme of the invention is as follows:
an HDAC inhibitor, wherein the inhibitor is a bifendate derivative shown as general formulas (I) - (IV) and a pharmaceutically acceptable salt or a deuteron thereof:
wherein, in the general formula (I) and (II), W is (CH)2)n1Or at least 1R1Substituted C6-14 aryl, C5-14 heteroaryl, C7-12 aralkyl, C6-12 heteroaralkyl;
in the general formula (III), X is at least 1R2Substituted C6-14 aryl, C5-14 heteroaryl, C7-12 aralkyl, C6-12 heteroaralkyl orWherein Y is at least 1R3Substituted C6-14 aryl, C5-14 heteroaryl;
in the general formula (IV), Z is at least substituted by 1R4Substituted C6-14 aryl, C5-14 heteroaryl;
n1＝1-9；R1、R2、R3、R4each independently selected from hydrogen, halogen, hydroxy, cyano, carbamoyl, C1-6 alkyl, C1-6 alkoxy, C2-6 unsaturated aliphatic hydrocarbon; r5Is hydrogen, C1-6 alkyl orn2＝1-4。
In a further embodiment, the bifendate derivatives represented by the general formulae (I) to (IV) are selected from the following compounds:
in a further embodiment, the HDAC inhibitor is a selective HDAC 6inhibitor, the selective HDAC 6inhibitor is a biphenyldicarboxylate derivative represented by the following general formulas (I) and (III) and a pharmaceutically acceptable salt or a deuteroxide thereof,
wherein in the general formula (I), W is at least 1R1Substituted C6-14 aryl, C5-14 heteroaryl; in the general formula (III), X is at least 1R2Substituted C6-14 aryl, C5-14 heteroaryl orAnd Y is at least 1R3Substituted C6-14 aryl, C5-14 heteroaryl; r1、R2、R3Each independently selected from hydrogen, halogen, hydroxy, cyano, carbamoyl, C1-6 alkyl, C1-6 alkoxy, C2-6 unsaturated aliphatic hydrocarbon; r5Is hydrogen, C1-6 alkyl orn2＝1-4。
Further, the HDAC inhibitor is a selective HDAC1/2/3 inhibitor, the HDAC1/2/3 inhibitor is a biphenyldicarboxylate derivative represented by the general formula (IV) and a pharmaceutically acceptable salt or a deuteron thereof,
wherein,in the general formula (IV), Z is at least substituted by 1R4Substituted C6-14 aryl, C5-14 heteroaryl; r4Independently selected from hydrogen, halogen, hydroxyl, cyano, carbamoyl, C1-6 alkyl, C1-6 alkoxy, C2-6 unsaturated aliphatic hydrocarbon.
The present invention also provides an HDAC inhibitor composition comprising the HDAC inhibitor described above and at least one pharmaceutically acceptable carrier or excipient.
In a further aspect, the HDAC inhibitor composition further comprises at least one additional therapeutic agent.
The dosage form of the HDAC inhibitor composition is any clinically or pharmaceutically acceptable dosage form.
The invention also provides application of the HDAC inhibitor in preparing medicaments for resisting tumors, autoimmune diseases, inflammations or Alzheimer's diseases.
In addition, the invention also provides an application of the pharmaceutical composition or the HDAC inhibitor composition in preparing medicaments for resisting tumors, autoimmune diseases, inflammations or Alzheimer's diseases.
The dosage of the compound of the invention is 1mg-1000 mg/day, and the dosage can be deviated from the range according to the severity of the disease or the dosage form.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs.
Wherein "halogen" means fluorine, chlorine, bromine or iodine.
"C6-14 aryl" refers to an all carbon monocyclic or fused polycyclic group of 6 to 14 carbon atoms having a completely conjugated pi-electron system. Non-limiting examples thereof are: benzene ring, naphthalene ring, anthracene ring.
"C5-14 heteroaryl" refers to a non-all carbon monocyclic or fused polycyclic group of 5 to 14 ring atoms with a fully conjugated pi-electron system. Non-limiting examples thereof are: pyridine, imidazole, thiophene, furan, thiazole, purine, indole and azaindole.
"C6-12 arylheteroalkyl" means a group having an alkyl group attached to an heteroaryl group containing 6 to 12 carbon atoms;
"C7-12 aralkyl" means a group having an alkyl group attached to an aryl group having 7 to 12 carbon atoms;
"C1-6 alkyl" refers to an alkyl group of 1to 6 carbon atoms;
"C1-6 alkoxy" means that one hydrogen atom of an alkane of 1to 6 carbon atoms is replaced by an oxygen atom. Such as methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, tert-butoxy, n-pentoxy, neopentoxy, n-hexoxy, and the like;
"C2-6 unsaturated aliphatic hydrocarbon group" refers to a straight or branched chain alkenyl or alkynyl group having 2 to 6 carbon atoms and containing a double bond or a triple bond. Non-limiting examples of unsaturated aliphatic hydrocarbon groups are: vinyl, 1-propenyl, 2-propenyl, ethynyl and the like.
The compound or the pharmaceutically acceptable salt or the deuteron thereof has the same effect, wherein the pharmaceutically acceptable salt is the salt of the general formula (I), (II), (III) or (IV), and the pharmaceutically acceptable salt is hydrochloride, sulfate, phosphate, maleate, fumarate, citrate, methanesulfonate, tartrate, sodium salt or potassium salt.
The "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable carrier, and includes diluents, excipients (e.g., water, etc.), fillers (e.g., starch, etc.), binders (e.g., cellulose derivatives, gelatin, etc.), humectants (e.g., glycerin, etc.), disintegrants (e.g., agar-agar, calcium carbonate, etc.), absorption enhancers (e.g., quaternary ammonium compounds, etc.), surfactants (e.g., cetyl alcohol, etc.), adsorbent carriers (e.g., kaolin, bentonite, etc.), lubricants (e.g., talc, etc.), and flavoring agents, sweeteners, etc., as necessary.
The "pharmaceutically acceptable dosage form" is suitable for administration by any suitable route, such as the oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) route. These formulations may be prepared by any method known in the art of pharmacy. For example, by admixing the active ingredient with a carrier or excipient.
The HDAC inhibitor is a bifendate derivative with HDAC inhibitory activity and a pharmaceutically acceptable salt or a deuterate thereof. The bifendate is a key intermediate for synthesizing Chinese medicine monomer schisandrin C, can be used as schisandrin C analogue structurally, is a common medicine for relieving transaminase rise caused by viral hepatitis and drug-induced liver injury in clinic, and has excellent safety. Recent studies have shown that biphenyldicarboxylate also has potential application value in tumor treatment, but studies on HDAC inhibitor discovery based on the basic skeleton of biphenyldicarboxylate have not been found yet.
Through multiple experiments, the bifendate derivatives prepared by the invention have HDAC (histone deacetylase) inhibition activity, and most compounds can inhibit HDAC1 or/and HDAC6 with high strength. Part of the compounds can inhibit HDAC1 at high intensity, show remarkable anti-tumor cell proliferation activity and have excellent selectivity on HDAC 1/2/3. Some other compounds show excellent HDAC6 selectivity while inhibiting HDAC6 at high intensity. Pharmacodynamic experiments show that the compound can be used as a therapeutic drug for tumors, autoimmune diseases, inflammations or Alzheimer's disease.
Detailed Description
Taking compound 1 (route I), compounds 2, 9, 12, 21 (route II) as examples, the compounds of the invention are prepared as follows:
alternatively, compounds 3-6 are prepared according to reference scheme I; compounds 7,8, 10, 11, 13-20 were prepared according to scheme II. It should be noted that the following specific examples are included for purposes of illustration and are not to be construed as limiting the scope of the invention. Further, it should be understood that various changes or modifications can be made by those skilled in the art after reading the contents of the present invention, and such equivalents also fall within the scope of the invention defined by the appended claims.
In route I: a is LiAlH4(lithium aluminum hydride), THF (tetrahydrofuran), 0 ℃;
b is MsCl (methanesulfonyl chloride), TEA (triethylamine), DCM (dichloromethane), rt (room temperature);
c is Methyl-8-aminoctanoate hydrochloride (8-aminocaprylic acid Methyl ester hydrochloride), TEA, CH3CN (acetonitrile), 40 ℃;
d is NH2OK (hydroxylamine potassium), CH3OH (methanol), rt.
In route II: a is Benzylamine (Benzylamine), TEA, CH3CN，40℃；
b is HCOONH4(ammonium formate), Pd/C (10%) (10% palladium on carbon), CH3OH,N2(nitrogen protection), 50 ℃;
c is methyl 4- (chloromethyl) benzoate (4-chloromethyl methyl benzoate), Cs2CO3(cesium carbonate), KI (potassium iodide), CH3CN，N2，80℃；
d is NH2OK，CH3OH，rt；
e is 5- (methoxycarbonyl) thiophene-2-carboxylic acid, EDC. HCl (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride), HOBT (1-hydroxybenzotriazole), DIPEA (N, N-diisopropylethylamine), DCM, rt;
f is methyl 4- (aminomethyl) benzoate hydrochloride, triphosgene, TEA, DCM, 0 deg.C-rt;
g is ethyl 2-chloropyrimidine-5-carboxylate (ethyl 2-chloropyrimidine-5-carboxylate), Cs2CO3KI (potassium iodide), CH3CN,N2，80℃。
Example 1:
synthesis of 8- (4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) -N-hydroxyoctanoylamide (1)
(1) Synthesis of (7,7' -dimethoxy- [4,4' -dibenzo [ d ] [1,3] dioxanone ] -5,5' -) dimethanol (23)
Biphenyl diester (10.0g,23.9mmol,1eq) was added to a round-bottomed flask, dissolved in anhydrous THF (150mL), and LiAlH was added in portions under ice bath4(2.72g,71.7mmol,3eq), after the addition was complete, the reaction was transferred to room temperature for 4 h. In ice bath, ice water was added to remove excess LiAlH4Suction filtration, DCM rinse, concentration of the filtrate, sequential use of saturated NaHCO3Washing the solution with water and saturated brine, anhydrous Na2SO4Drying, filtering, and concentrating the filtrate under reduced pressure to obtain white solid 8.33 g.
(2) Synthesis of (7,7' -dimethoxy- [4,4' -dibenzo [ d ] [1,3] dioxanone ] -5,5' -) bis (methylene) dimesylate (24)
Intermediate 23(5.50g,15.2mmol,1eq) was added to a round bottom flask, dissolved in DCM (50mL), and TEA (6.33mL,45.5mmol,3eq) and MsCl (3.84mL,45.5mmol,3eq) were added sequentially under ice bath and transferred to room temperature for 8 h. The system is sequentially saturated with NaHCO3Washing the solution with water and saturated brine, anhydrous Na2SO4Drying, filtering and concentrating the filtrate under reduced pressure to obtain 7.57g of yellow oily substance which is directly used for the next reaction.
(3) Synthesis of methyl 8- (4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) octanoate (25)
A round bottom flask was charged with intermediate 24(200mg,0.386mmol,1eq), methyl 8-aminocaprylate hydrochloride (323mg,1.54mmol,4eq), TEA (0.322mL,2.31mmol,6eq), and CH in that order3CN (3mL), reacted at 40 ℃ for 8 h. Rotate to remove CH3CN, in turn with saturated NaHCO3The solution, water and saturated brine are washed, DCM is used for extraction, and anhydrous Na is added2SO4Drying, filtering, concentrating the filtrate under reduced pressure, and performing silica gel column chromatography to obtain light yellow solid 163 mg.
(4) Synthesis of 8- (4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) -N-hydroxyoctanoyl amide (1)
NH2Preparation of OK methanol solution: KOH (9.70g,173mmol,1.5eq) was weighed into a round bottom flask and dissolved in methanol (24 mL); weighing NH2OH & HCl (8.00g,115mmol,1eq) was placed in a three-necked flask and dissolved in methanol (40 mL); under ice bath, KOH methanol solution was added dropwise to NH2OH HCl in methanol solution, N2Reacting for 1h under protection, and performing suction filtration to obtain filtrate for later use.
In a round bottom flask, intermediate 25(130mg, 0.293mmol,1eq), NH were added sequentially2OK methanol solution (4mL) was reacted at room temperature for 4 h. Adjusting pH to 7 with 1N HCl solution, separating out solid, vacuum filtering, washing filter cake with water, drying, and performing silica gel column chromatography to obtain yellow solid 80 mg;1H NMR(400MHz,DMSO-d6):δ10.38(s,1H),8.67(s,1H),6.98(s,2H),6.12(s,2H),6.07(s,2H),4.41-4.28(m,1H),4.14-4.00(m,1H),3.90(s,6H),3.78-3.66(m,1H),3.22-2.85(m,3H),1.96(t,J＝7.2Hz,2H),1.86-1.76(m,1H),1.75-1.65(m,1H),1.51(t,J＝6.8Hz,2H),1.37-1.27(m,6H)；ESI-MS:501.19[M+H]+。
example 2:
synthesis of 4- ((4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) methyl) -N-hydroxybenzamide (2)
(1) Synthesis of 7-benzyl-4, 10-dimethoxy-7, 8-dihydro-6H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine (25)
A round bottom flask was charged with intermediate 24(7.57g,14.6mmol,1eq), CH in that order3CN (50mL), benzylamine (6.38mL,58.4mmol,4eq) and TEA (8.12mL,58.4mmol,4eq) were reacted at 40 ℃ for 8 h. Rotate to remove CH3CN, in turn with saturated NaHCO3The solution, water and saturated brine are washed, DCM is used for extraction, and anhydrous Na is added2SO4Drying, filtering, decompressing and concentrating the filtrate, and recrystallizing with ethyl acetate to obtain 5.01g of white solid.
(2) Synthesis of 4, 10-dimethoxy-7, 8-dihydro-6H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine (26)
A round bottom flask was charged with intermediate 25(5.00g,11.5mmol,1eq), HCOONH in that order4(4.00g,63.4mmol,5.5eq), 10% Pd/C (1.10g), methanol (50mL), N2Protecting, and stirring and reacting for 10h at 50 ℃. Filtering to remove Pd/C, rinsing with methanol, concentrating the filtrate under reduced pressure, sequentially adding saturated NaHCO3The solution, water and saturated brine are washed, DCM is used for extraction, and anhydrous Na is added2SO4Drying, filtering, concentrating the filtrate under reduced pressure, and performing silica gel column chromatography to obtain light yellow solid 2.09 g.
(3) Synthesis of methyl 4- ((4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) methyl) benzoate (27)
The intermediate 26(150mg,0.437mmol,1eq), methyl 4-chloromethylbenzoate (96.8mg,0.524mmol,1.2eq), and Cs were added to a round-bottomed flask in this order2CO3(185mg,0.524mmol,1.2eq), KI (87mg,0.524mmol,1.2eq) and anhydrous CH3CN(2mL)，N2Protecting, and reacting at 80 ℃ for 9 h. Rotary wrenchRemoving CH3CN, in turn with saturated NaHCO3The solution, water and saturated brine are washed, DCM is used for extraction, and anhydrous Na is added2SO4Drying, filtering, concentrating the filtrate under reduced pressure, and performing silica gel column chromatography to obtain yellow solid 206 mg.
(4) Synthesis of 4- ((4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) methyl) -N-hydroxybenzamide (2)
Compound 2 was prepared according to example 1, step (4).
1H NMR(400MHz,DMSO-d6):δ11.20(s,1H),9.03(s,1H),7.76(d,J＝8.0Hz,2H),7.48(d,J＝8.0Hz,2H),6.65(s,2H),6.05(s,2H),6.00(s,2H),3.88(s,6H),3.72(d,J＝13.6Hz,1H),3.54(d,J＝13.6Hz,1H),3.44(d,J＝12.8Hz,2H),3.03(d,J＝12.4Hz,2H)；ESI-MS:493.24[M+H]+。
Example 3:
synthesis of 5- (4, 10-dimethoxy-7, 8-dihydro-6H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carbonyl) -N-hydroxythiophene-2-carboxamide (9)
(1) Synthesis of methyl 5- (4, 10-dimethoxy-7, 8-dihydro-6H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carbonyl) thiophene-2-carboxylate (28)
5-methoxycarbonylthiophene-2-carboxylic acid (89.5mg,0.481mmol,1.1eq), EDC & HCl (92.1mg,0.481mmol,1.1eq), HOBt (65.4mg,0.481mmol,1.1eq), DIPEA (217. mu.L, 1.31mmol,3eq) and DCM (2mL) were added in this order to a round-bottomed flask and stirred at room temperature for 1 h. Intermediate 26(150mg,0.437mmol,1eq) was added and the reaction was continued at room temperature for 4 h. Sequentially with saturated NaHCO3The solution, water and saturated brine are washed, DCM is used for extraction, and anhydrous Na is added2SO4DryingFiltering, concentrating the filtrate under reduced pressure, and performing silica gel column chromatography to obtain yellow solid 208 mg.
(2) Synthesis of 5- (4, 10-dimethoxy-7, 8-dihydro-6H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carbonyl) -N-hydroxythiophene-2-carboxamide (9)
Compound 9 was prepared according to step (4) in example 1.
1H NMR(400MHz,DMSO-d6):δ11.41(s,1H),9.29(s,1H),7.59(d,2.8Hz,1H),7.53(d,4.0Hz,1H),6.88(s,1H),6.75(s,1H),6.09(s,2H),6.04(s,2H),5.17-4.71(m,2H),4.27-4.01(m,1H),3.85(s,6H),3.65-3.49(m,1H)；ESI-MS:513.08[M+H]+。
Example 4:
synthesis of N- (4- (hydroxycarbamoyl) benzyl) -4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (12)
(1) Synthesis of methyl 4- ((4, 10-dimethoxy-7, 8-dihydro-6H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamido) methyl) benzoate (29)
Methyl 4-aminomethylbenzoate hydrochloride (150mg,0.744mmol,1eq) was added to a round-bottom flask, dissolved in a DCM/saturated sodium bicarbonate solution system (V: V ═ 1:1,4mL), triphosgene (73.6mg,0.248mmol,0.33eq) was added under ice-bath, and the reaction was stirred for 0.5 h. Intermediate 26(255mg,0.744mmol,1eq) and TEA (310. mu.L, 2.23mmol,3eq) were added and transferred to room temperature for 6 h. Sequentially with saturated NaHCO3The solution, water and saturated brine are washed, DCM is used for extraction, and anhydrous Na is added2SO4Drying, filtering, concentrating the filtrate under reduced pressure, and performing silica gel column chromatography to obtain 145mg of yellow solid.
(2) Synthesis of N- (4- (hydroxycarbamoyl) benzyl) -4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (12)
Compound 12 was prepared according to step (4) in example 1.
1H NMR(400MHz,DMSO-d6):δ11.18(s,1H),9.01(s,1H),7.70(d,8.0Hz,2H),7.35(d,8.0Hz,2H),7.22(t,4.8Hz,1H),6.78(s,2H),6.07(s,2H),6.02(s,2H),4.74(d,J＝13.2Hz,2H),4.36(dd,J＝15.2,4.8Hz,1H),4.28(dd,J＝15.2,4.8Hz,1H),3.87(s,6H),3.47(d,J＝13.2Hz,2H)；ESI-MS:536.12[M+H]+。
Example 5:
synthesis of 2- (4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) -N-hydroxypyrimidine-5-carboxamide (21)
(1) Synthesis of ethyl 2- (4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) pyrimidine-5-carboxylate (30)
Intermediate 30 was prepared according to example 2, step (3), substituting only methyl 4-chloromethylbenzoate with ethyl 2-chloro-5-pyrimidinecarboxylate.
(2) Synthesis of 2- (4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepin-7-yl) -N-hydroxypyrimidine-5-carboxamide (21)
Compound 21 was prepared according to step (4) of reference example 1.
1H NMR(400MHz,DMSO)δ11.13(s,1H),9.04(s,1H),8.76(s,2H),6.81(s,2H),6.08(s,2H),6.03(s,2H),5.43(d,J＝13.6Hz,2H),3.85(s,6H),3.67(d,J＝13.6Hz,2H)；ESI-MS:481.10[M+H]+。
Example 6:
synthesis of 5- ((4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) methyl) -N-hydroxythiophene-2-carboxamide (3)
Compound 3 was prepared according to example 5, replacing only ethyl 2-chloro-5-pyrimidinecarboxylate with methyl 5- (bromomethyl) thiophene-2-carboxylate.
1H NMR(400MHz,DMSO-d6):δ11.15(s,1H),9.09(s,1H),7.50(s,1H),7.08(s,1H),6.67(s,2H),6.05(s,2H),6.01(s,2H),4.12-4.01(m,1H),3.89(s,6H),3.70(d,12.4Hz,1H),3.54(d,11.6Hz,2H),3.04(d,11.2Hz,2H)；ESI-MS:498.94[M+H]+。
Example 7:
synthesis of 5- ((4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) methyl) -N-hydroxyfuran-2-carboxamide (4)
Compound 4 was prepared according to example 5, substituting only ethyl 2-chloro-5-pyrimidinecarboxylate with ethyl 5- (chloromethyl) furan-2-carboxylate.
1H NMR(400MHz,DMSO-d6):δ11.11(s,1H),9.04(s,1H),7.02(d,2.8Hz,1H),6.72(s,2H),6.55(d,J＝3.2Hz,1H),6.04(s,2H),6.00(s,2H),3.88(s,6H),3.69(d,J＝14.0Hz,1H),3.52(d,J＝14.4Hz,1H),3.47(d,J＝12.8Hz,2H),3.04(d,J＝12.4Hz,2H)；ESI-HRMS:483.1338[M+H]+。
Example 8:
synthesis of 2- ((4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) methyl) -N-hydroxypyrimidine-5-carboxamide (5)
Compound 5 was prepared according to example 5, substituting only ethyl 2-chloro-5-pyrimidinecarboxylate with methyl 2- (bromomethyl) pyrimidine-5-carboxylate. ESI-MS 495.15[ M + H ]]+。
Example 9:
synthesis of 6- ((4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) methyl) -N-hydroxynicotinamide (6)
Compound 6 was prepared according to example 5, substituting only ethyl 2-chloro-5-pyrimidinecarboxylate with methyl 2- (bromomethyl) pyridine-5-carboxylate.
1H NMR(400MHz,DMSO-d6):δ11.35(s,1H),9.16(s,1H),8.87(d,J＝1.6Hz,1H),8.13(dd,J＝8.0,1.6Hz,1H),7.64(d,J＝8.0Hz,1H),6.71(s,2H),6.05(s,2H),6.01(s,2H),3.93-3.83(m,7H),3.65(d,J＝14.0Hz,1H),3.48(d,J＝12.4Hz,2H),3.09(d,J＝12.4Hz,2H)；ESI-HRMS:494.1547[M+H]+。
Example 10:
synthesis of 8- (4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) -N-hydroxy-8-oxooctanoyl amide (7)
Compound 7 was prepared according to example 3, substituting only 5-methoxycarbonylthiophene-2-carboxylic acid with monomethyl suberate.
1H NMR(400MHz,DMSO-d6):δ10.33(s,1H),8.66(s,1H),6.97(s,1H),6.75(s,1H),6.06(s,2H),6.01(s,2H),5.11(d,J＝13.6Hz,1H),4.65(d,J＝13.2Hz,1H),3.88(s,3H),3.86(s,3H),3.72(d,J＝13.2Hz,1H),3.27(d,J＝13.6Hz,1H),2.58-2.54(m,1H),2.44-2.37(m,1H),1.94(t,J＝7.2Hz,2H),1.56-1.46(m,4H),1.35-1.27(m,4H)；ESI-MS:515.09[M+H]+。
Example 11:
synthesis of 4- (4, 10-dimethoxy-7, 8-dihydro-6H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carbonyl) -N-hydroxybenzamide (8)
Compound 8 was prepared according to example 3, substituting only 5-methoxycarbonylthiophene-2-carboxylic acid with monomethyl terephthalate.
1H NMR(400MHz,DMSO-d6):δ11.34(s,1H),9.16(s,1H),7.87(d,J＝8.0Hz,2H),7.56(d,J＝8.0Hz,2H),6.95(s,1H),6.41(s,1H),6.08(s,2H),6.03(s,2H)5.11(d,J＝13.6Hz,1H),4.27(d,J＝13.2Hz,1H),3.98-3.85(m,4H),3.81(s,3H),3.58(d,J＝13.6Hz,1H)；ESI-MS:506.99[M+H]+。
Example 12:
synthesis of 5- (4, 10-dimethoxy-7, 8-dihydro-6H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carbonyl) -N-hydroxyfuran-2-carboxamide (10)
Compound 10 was prepared according to example 3, substituting only 5-methoxycarbonylthiophene-2-carboxylic acid with 5-methoxycarbonylfuran-2-carboxylic acid. ESI-MS 497.11[ M + H ]]+。
Example 13:
synthesis of 4- (2- (4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-) -2-oxoethyl) -N-hydroxybenzamide (11)
Compound 11 was prepared according to example 3, substituting only 5-methoxycarbonylthiophene-2-carboxylic acid with 4- (methoxycarbonyl) phenylacetic acid. ESI-MS 521.16[ M + H ]]+。
Example 14:
synthesis of N- ((5- (hydroxycarbamoyl) pyrimidin-2-) methyl) -4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (13)
Compound 13 was prepared according to example 4, substituting only methyl 4-aminomethylbenzoate hydrochloride with methyl 2-aminomethylpyrimidine-5-carboxylate hydrochloride. ESI-MS 538.16[ M + H ]]+。
Example 15:
synthesis of N- ((5- (hydroxycarbamoyl) thiophen-2-) methyl) -4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (14)
Compound 14 was prepared according to example 4, substituting only 4-aminomethylbenzoic acid methyl ester hydrochloride with 5-aminomethylthiophene-2-carboxylic acid methyl ester hydrochloride.
1H NMR(400MHz,DMSO-d6):δ10.35(s,1H),9.10(s,0.7H),8.55(s,0.3H),7.51-7.40(m,1H),7.34(t,5.2Hz,1H),6.95(d,J＝2.8Hz,1H),6.78(s,2H),6.07(s,2H),6.02(s,2H),4.71(d,J＝13.2Hz,2H),4.48(dd,J＝15.6,5.2Hz,3H),4.37(dd,J＝15.6,5.2Hz,3H),3.88(s，6H)，3.46(d,J＝13.2Hz,2H)；ESI-HRMS:542.1226[M+H]+。
Example 16:
(E) synthesis of (E) -N- (4- (3- (hydroxyamino) -3-oxoprop-1-en-1-) benzyl) -4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (15)
Compound 15 was prepared according to example 4, substituting only methyl 4-aminomethylbenzoate hydrochloride with methyl (E) -4-aminomethylbenzoate hydrochloride.
1H NMR(400MHz,DMSO-d6):δ10.74(s,1H),9.03(s,1H),7.51(d,J＝7.6Hz,2H),7.44(d,J＝15.6Hz,1H),7.33(d,J＝8.0Hz,2H),7.18(t,J＝5.2Hz,1H),6.78(s,2H),6.44(d,J＝15.6Hz,1H),6.07(s,2H),6.02(s,2H),4.74(d,J＝13.2Hz,2H),4.36(dd,J＝15.6,5.2Hz,1H),4.25(dd,J＝15.6,5.2Hz,1H),3.87(s,6H),3.47(d,J＝13.2Hz,2H)；ESI-HRMS:562.1818[M+H]+。
Example 17:
synthesis of N- (4- (hydroxycarbamoyl) benzyl) -4, 10-dimethoxy-N-methyl-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (16)
Compound 16 was prepared according to example 4, substituting only methyl 4-aminomethylbenzoate hydrochloride with methyl 4- ((methylamino) methyl) benzoate hydrochloride.
1H NMR(400MHz,DMSO-d6):δ11.20(s,1H),9.02(s,1H),7.76(d,J＝7.6Hz,2H),7.42(d,J＝7.2Hz,2H),6.75(s,2H),6.06(s,2H),6.01(s,2H),4.53(d,J＝15.2Hz,1H),4.28(d,J＝10.8Hz,1H),4.23(d,J＝12.8Hz,2H),3.87(s,6H),3.61(d,J＝12.8Hz,2H),2.79(s,3H)；ESI-MS:550.18[M+H]+。
Example 18:
synthesis of N- (4- (hydroxycarbamoyl) benzyl) -N-ethyl-4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (17)
Compound 17 was prepared according to example 4, substituting only methyl 4-aminomethylbenzoate hydrochloride with methyl 4- ((ethylamino) methyl) benzoate hydrochloride.1H NMR(400MHz,DMSO-d6):δ11.20(s,1H),9.02(s,1H),7.75(d,J＝8.0Hz,2H),7.43(d,J＝7.6Hz,2H),6.73(s,2H),6.07(s,2H),6.01(s,2H),4.56(d,J＝15.6Hz,1H),4.32-4.17(m,3H),3.88(s,6H),3.62(d,J＝12.8Hz,2H),3.26–3.19(m,1H),3.16–3.04(m,1H),1.14(t,J＝6.4Hz,3H)；ESI-HRMS:564.1976[M+H]+。
Example 19:
synthesis of N- (4- (hydroxycarbamoyl) benzyl) -N-N-butyl-4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (18)
Compound 18 was prepared according to example 4, substituting only methyl 4-aminomethylbenzoate hydrochloride with methyl 4- ((butylamino) methyl) benzoate hydrochloride.
1H NMR(400MHz,DMSO-d6):δ11.21(s,1H),9.04(s,1H),7.74(d,J＝8.0Hz,2H),7.41(d,J＝8.0Hz,2H),6.69(s,2H),6.07(s,2H),6.01(s,2H),4.59(d,J＝15.6Hz,1H),4.24(d,J＝13.2Hz,2H),4.17(d,J＝16.0Hz,1H),3.87(s,6H),3.62(d,J＝13.2Hz,2H),3.28-3.20(m,1H),3.01-2.91(m,1H),1.61-1.51(m,2H),1.32-1.27(m,2H),0.87(t,J＝10.0Hz,3H)；ESI-MS:592.22[M+H]+。
Example 20:
synthesis of N- (4- (hydroxycarbamoyl) benzyl) -N- (3-hydroxypropyl) -4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (19)
Compound 19 was prepared according to example 4, substituting only methyl 4-aminomethylbenzoate hydrochloride with methyl 4- (((3-hydroxypropyl) amino) methyl) benzoate hydrochloride.
1H NMR(400MHz,DMSO-d6):δ11.20(s,1H),9.03(s,1H),7.74(d,J＝8.4Hz,2H),7.39(d,J＝8.4Hz,2H),6.73(s,2H),6.06(s,2H),6.01(s,2H),4.58(d,J＝15.6Hz,1H),4.49(t,J＝5.2Hz,1H),4.29-4.19(m,3H),3.87(s,6H),3.61(d,J＝13.2Hz,2H),3.45-3.42(m,2H),3.30-3.23(m,1H),3.10-3.01(m,1H),1.79-1.70(m,2H)；ESI-HRMS:594.2084[M+H]+。
Example 21:
(E) synthesis of (E) -N- (4- (3- (hydroxyamino) -3-oxoprop-1-en-1-yl) benzyl) -N-methyl-4, 10-dimethoxy-6, 8-dihydro-7H- [1,3] dioxa [4',5':3,4] benzo [1,2-c ] [1,3] dioxa [4',5':5,6] benzo [1,2-e ] azepine-7-carboxamide (20)
Compound 20 was prepared according to example 4, substituting only methyl 4-aminomethylbenzoate hydrochloride with methyl (E) -3- (4- ((methylamino) methyl) phenyl) acrylate hydrochloride. ESI-HRMS 576.1980[ M + H ]]+。
Example 22: HDAC1 enzyme inhibitory Activity of Compounds of the invention
In this example, a broad-spectrum HDACs inhibitor SAHA from the market was used as a positive control, and fluorescence-based HDAC Activity Assay was used to evaluate the HDAC1 enzyme inhibitory Activity of the compound of the present invention and SAHA. Other compounds of the present invention have similar beneficial effects to those listed below, but this should not be understood as the only beneficial effects of the compounds of the present invention.
The HDAC1 enzyme inhibitory activity was tested by the following steps: preparing a DMSO solution of a compound to be detected, and sequentially preparing a buffer solution, an enzyme solution and a corresponding Substrate/Trypsin mixed solution according to the kit instructions; mixing the compound solution with gradient concentration, enzyme solution and hydrate/TrypsinAdding the solution into 384-well plates respectively to prepare a catalytic reaction system (no compound control and no enzyme control wells); after incubation for a certain time at room temperature, continuously reading a fluorescence signal value by using a Synergy enzyme-labeling instrument, selecting a linear reaction section to obtain a slope (slope), and further calculating the inhibition rate and IC (integrated circuit) under each concentration50Obtained by fitting GraphPad Prism 5 software.
Table 1: inhibitory Activity of Compounds on HDAC1 enzymes
In the above table: "+ ++" represents 0-20 nM; "+ + + +" represents 20-100 nM.
As can be seen from the data in table 1, all of the compounds listed exhibited significant HDAC1 inhibitory activity, which was comparable to or superior to SAHA.
Example 22: antitumor cell proliferation Activity of Compounds of the present invention
In this example, SAHA was used as a positive control, and the anti-proliferative activity of the compound of the present invention and SAHA on lung cancer cell line A549 and colon cancer cell line HCT116 was evaluated by CCK-8 method. Other compounds of the present invention have similar beneficial effects to those listed below, but this should not be understood as the only beneficial effects of the compounds of the present invention.
The test procedure for the anti-tumor cell proliferation activity was: digesting and collecting tumor cells, inoculating the cells in a 96-well culture plate at a certain density, and placing the plate in an incubator (37 ℃, 5% CO)2) Overnight. After the compound is applied for 72h, the culture medium is discarded, and the cells are washed gently with PBS 3 times. Subsequently, a certain volume of medium and CCK-8 were added to each well of the plate, and the incubation was continued for a certain period of time. Finally, a multifunctional microplate reader is adopted to measure the OD value of absorbance under the wavelength of 570nm, and the inhibition rate, GI is calculated50Values were fitted by GraphPad Prism 5 software.
Table 2: antitumor cell proliferation activity of the compound
In the above table: "+ ++" represents < 0.1. mu.M; "+ + + +" represents 0.1-1.0. mu.M; "+ +" represents 1.0-10. mu.M; "+" represents 10-50. mu.M.
As can be seen from the data in Table 2, all the compounds listed exhibit significant anti-tumor cell proliferation activity, which is comparable to or superior to SAHA.
Example 23: HDAC6 Selectivity or class I HDACs Selectivity of Compounds of the invention
The literature indicates that the selectivity of the compounds for inhibiting HDAC isoforms can be reflected by the activity of inhibiting HDAC1, 2, 3, 6,8, 10, 11 enzymes (Yu CW, et al Quinazolin-2,4-dione-Based Hydroxamic Acids as selective Histone deacylase-6 Inhibitors for Treatment of Non-Small cell Lung cancer. J Med Chem,2019,62)
Test methods for inhibitory activity of compounds against HDAC 6and other HDAC isomerases the catalytic reaction system substrates were only replaced when the corresponding enzyme inhibitory activity was tested, in reference to the test method for HDAC1 inhibitory activity.
The selectivity of the partial compounds of the present invention for HDAC6 or HDAC1/2/3 is further illustrated by the inhibitory activity data of HDAC1, 2, 3, 6,8, 10, 11. It is not to be understood that the present invention is only the following compounds which have selectivity for HDAC6 or HDAC 1/2/3.
Table 3: inhibitory Activity of Compounds on HDAC1, 2, 3, 6,8, 10, 11
In the above table: "+ ++" represents 0-10 nM; "+ + + +" represents 10-100 nM; "+ +" represents 100 and 1000 nM; "+" represents 1000-10000 nM.
As shown in table 3, compound 3 according to formula (I) and compounds 12, 19, and 20 according to formula (III) have significant inhibitory activity against HDAC6, but relatively weak inhibitory activity against HDACs 1,2, 3, 8, 10, and 11, and are selective HDAC6 inhibitors. The compound 21 has remarkable inhibiting activity on HDAC1, 2 and 3, and relatively weaker inhibiting activity on HDAC6, 8, 10 and 11, so the compound is a selective HDAC1/2/3 inhibitor. Compared with the broad-spectrum HDACs inhibitor SAHA, the compounds 3, 12, 19, 20 and 21 of the present invention, while significantly inhibiting the corresponding HDAC subtype, are beneficial to reduce the toxicity of SAHA to all HDAC subtype inhibition.
In conclusion, the compound prepared by the invention has good application prospect.

Claims (8)

1. A class of HDAC inhibitors characterized by: the inhibitor is a bifendate derivative shown in the following general formulas (I) to (IV) and a pharmaceutically acceptable salt or a deuteron thereof:
in the above formula, W is (CH)2)n1Or at least 1R1Substituted C6-14 aryl, C5-14 heteroaryl, C7-12 aralkyl, C6-12 heteroaralkyl;
x is at least 1R2Substituted C6-14 aryl, C5-14 heteroaryl, C7-12 aralkyl, C6-12 heteroaralkyl orWherein Y is at least 1R3Substituted C6-14 aryl, C5-14 heteroaryl;
z is at least 1R4Substituted C6-14 aryl, C5-14 heteroaryl;
n1＝1-9；R1、R2、R3、R4each independently selected from hydrogen, halogen, hydroxy, cyano, carbamoyl, C1-6 alkyl, C1-6 alkoxy, C2-6 unsaturated aliphatic hydrocarbon; r5Is hydrogen, C1-6 alkyl orn2＝1-4。
2. The class of HDAC inhibitors according to claim 1, wherein: the bifendate derivative is selected from the following compounds:
3. the class of HDAC inhibitors according to claim 1, wherein: the HDAC inhibitor is a selective HDAC 6inhibitor, the selective HDAC 6inhibitor is a bifendate derivative shown as general formulas (I) and (III) and a pharmaceutically acceptable salt or deuteron thereof,
wherein in the general formula (I), W is at least 1R1Substituted C6-14 aryl, C5-14 heteroaryl;
in the general formula (III), X is at least 1R2Substituted C6-14 aryl, C5-14 heteroaryl orAnd Y is at least 1R3Substituted C6-14 aryl, C5-14 heteroaryl; r1、R2、R3Each independently selected from hydrogen, halogen, hydroxy, cyano, carbamoyl, C1-6 alkyl, C1-6 alkoxy, C2-6 unsaturated aliphatic hydrocarbon; r5Is hydrogen, C1-6 alkyl orn2＝1-4。
4. The class of HDAC inhibitors according to claim 1, wherein: the HDAC inhibitor is a selective HDAC1/2/3 inhibitor, the HDAC1/2/3 inhibitor is a biphenyl diester derivative shown as a general formula (IV) and a pharmaceutically acceptable salt or a deuteron thereof,
wherein in the general formula (IV), Z is at least substituted by 1R4Substituted C6-14 aryl, C5-14 heteroaryl; r4Independently selected from hydrogen, halogen, hydroxyl, cyano, carbamoyl, C1-6 alkyl, C1-6 alkoxy, C2-6 unsaturated aliphatic hydrocarbon.
5. An HDAC inhibitor composition characterized by: comprising an HDAC inhibitor as claimed in any one of claims 1to 4 together with at least one pharmaceutically acceptable carrier or excipient.
6. The HDAC inhibitor composition of claim 5, wherein: further comprising at least one other therapeutic agent; the dosage form of the HDAC inhibitor composition is any clinically or pharmaceutically acceptable dosage form.
7. The use of an HDAC class inhibitor as claimed in any of claims 1-4 wherein: the HDAC inhibitor is applied to the preparation of anti-tumor, autoimmune disease, inflammation or Alzheimer disease drugs.
8. Use of a composition according to claim 5 or 6, characterized in that: the composition is applied to the preparation of anti-tumor, autoimmune disease, inflammation or Alzheimer's disease drugs.
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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111848629A (en) * 2020-07-15 2020-10-30 安徽中医药大学 mTOR/HDAC dual inhibitor and application thereof
CN113754627A (en) * 2021-09-03 2021-12-07 西北师范大学白银师科创新研究院 Preparation method of biphenylol acid
CN114805375A (en) * 2022-04-12 2022-07-29 徐州医科大学 N-phenyl alkoxy dibenzoazepine compound, preparation method and medical application thereof
WO2022203429A1 (en) * 2021-03-26 2022-09-29 Chong Kun Dang Pharmaceutical Corp. Composition for preventing or treating multiple sclerosis

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04202153A (en) * 1990-11-29 1992-07-22 Tokyo Tanabe Co Ltd New tricyclic composition having treating action on hepatic disease
CN102532150A (en) * 2012-02-27 2012-07-04 中国药科大学 Alkoxyl dibenzoazepine compound, and preparation method and medical application thereof
CN107266461A (en) * 2017-06-20 2017-10-20 徐州医科大学 A kind of alkoxy dibenzazepines class compound, its preparation method and medical usage

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04202153A (en) * 1990-11-29 1992-07-22 Tokyo Tanabe Co Ltd New tricyclic composition having treating action on hepatic disease
CN102532150A (en) * 2012-02-27 2012-07-04 中国药科大学 Alkoxyl dibenzoazepine compound, and preparation method and medical application thereof
CN107266461A (en) * 2017-06-20 2017-10-20 徐州医科大学 A kind of alkoxy dibenzazepines class compound, its preparation method and medical usage

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
XIAOKE GU 等: "Synthesis and biological evaluation of bifendate derivatives bearing 6,7-dihydro-dibenzo[c,e]azepine scaffold as potential P-glycoprotein and tumor metastasis inhibitors", 《EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY》 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111848629A (en) * 2020-07-15 2020-10-30 安徽中医药大学 mTOR/HDAC dual inhibitor and application thereof
WO2022203429A1 (en) * 2021-03-26 2022-09-29 Chong Kun Dang Pharmaceutical Corp. Composition for preventing or treating multiple sclerosis
CN113754627A (en) * 2021-09-03 2021-12-07 西北师范大学白银师科创新研究院 Preparation method of biphenylol acid
CN114805375A (en) * 2022-04-12 2022-07-29 徐州医科大学 N-phenyl alkoxy dibenzoazepine compound, preparation method and medical application thereof
CN114805375B (en) * 2022-04-12 2023-07-25 徐州医科大学 N-phenylalkoxy dibenzoazepine compound, preparation method and medical application thereof

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