Chimeric Molecules with Antitumor Activity, Their Preparation Methods and Applications

Abstract

The present invention discloses a chimeric molecule, a preparation method thereof, and application thereof in treating cancer, and particularly relates to a chimeric molecule that has an inhibitory effect on the PARP-1 target.

Description

Technical Field

[0001] The present invention relates to the field of cancer drugs, and discloses a chimeric molecule and a preparation method thereof, as well as the use of the compound and its pharmaceutical composition in treating cancer. Specifically, it relates to a chimeric molecule that has an inhibitory effect on the PARP-1 target. Background Art

[0002] There are three basic levels of protein regulation: the first is at the DNA level, where knockout knockout inactivates the DNA expressing the target protein; the second is at the mRNA level, where mRNA binding and inactivation of the target protein inhibits protein expression; and the third is at the protein level, where modifications such as methylation modulate the target protein's activity. The chimeric molecules involved in this invention are used to treat diseases by regulating the degradation of the target protein at the protein level.

[0003] PARP-1 chimeric molecules are bifunctional molecules composed of an E3 ubiquitin ligase recognition group, a target protein (PARP-1) recognition group, and a linker group. E3 ubiquitin ligases (hundreds of which are known in humans) confer specificity to their substrates for ubiquitination, and their specificity for certain protein substrates makes them more attractive than general proteasome inhibitors. Prior art suggests that an E3 ligase with therapeutic potential is Cereblon, a protein encoded by the human CB gene. CB homologs are highly conserved, indicating its physiological importance. Cereblon forms an E3 ubiquitin ligase complex with damaged DNA binding protein 1 (DDB1), Cullin-4A (CUL4A), and regulator of Cullin-1 (ROCI). This complex ubiquitinates a range of proteins, but the specific mechanisms remain unclear. Cereblon ubiquitination of target proteins leads to increased levels of fibroblast growth factor 8 (FGFB) and fibroblast growth factor 10 (FGF10), indicating the importance of this ubiquitinase complex in embryonic limb growth. Poly(ADP-ribose) polymerases (PARPs) are a class of nuclear enzymes widely found in eukaryotic cells, mediating DNA repair and maintaining genomic integrity. To date, studies have identified at least 18 members of the PARP enzyme family, with PARP-1 being the most abundant isoform. It accounts for over 90% of the intracellular ADP-ribosylation process and is highly evolutionarily conserved across eukaryotes, making it the most intensively studied. PARP-1 participates in the base excision repair pathway of single-stranded DNA damage by mediating the polymerization of adenosine diphosphate-ribose (ADP-ribose) and transferring it from nicotinamide adenine dinucleotide (NAD+) to receptor proteins. To maintain normal physiological function, cells must possess multiple DNA damage detection and repair mechanisms to ensure timely and accurate repair of damaged DNA. Many current anticancer drugs target tumors by damaging the DNA of tumor cells. However, tumor cells can activate their own DNA damage repair mechanisms, leading to resistance to these therapies. Therefore, an important approach to cancer treatment is to block the DNA repair pathway. PARP-1 has become an effective anti-tumor target. The goal of chimeric molecules is to bind to the target protein PARP-1 through PARP-1 recognition groups, especially PARP-1 inhibitors. The E3 ubiquitin ligase recognition group leads to ubiquitination of the target protein, ultimately leading to its degradation by the proteasome.

[0004] From knocking out functional genes at the genetic level to interfering with mRNA levels, there are many ways to study protein expression and regulation. Chimeric molecules can directly regulate protein expression at the protein level without affecting the expression of DNA and mRNA, thus avoiding unpredictable effects at the gene and RNA levels. At the same time, chimeric molecules target protein degradation and have better anti-tumor effects than PARP-1 inhibitors. They may be developed into drugs and become new treatment methods. Summary of the Invention

[0005] The present invention aims to provide novel chimeric molecules or pharmaceutically acceptable salts, hydrates, or prodrugs thereof. These compounds have the function of inducing PARP-1 degradation and can be used to prepare novel anti-tumor drugs. The tumor may be, but is not limited to, multiple myeloma, gastric cancer, lung cancer, breast cancer, esophageal cancer, colon cancer, medulloblastoma, acute myeloid leukemia, chronic leukemia, prostate cancer, hepatoma, renal cell tumor, cervical cancer, skin cancer, ovarian cancer, colon cancer, glioma, thyroid cancer, or pancreatic cancer. The present invention also aims to provide a method for synthesizing the novel chimeric molecules. Another object of the present invention is to provide a pharmaceutical preparation containing the novel chimeric molecules.

[0006] The present invention first provides a compound as shown in Formula I or a pharmaceutically acceptable salt thereof:

[0007] ALB(I)

[0008] Wherein: L is a linking group, and one atom on A and B is replaced by one end of the linking group L;

[0009] A is shown in Formula II:

[0010]

[0011] in:

[0012] (R) n Where R is halogen or C1-C8 alkyl, and n is an integer selected from 0-2;

[0013] R 1 is hydrogen or C1-C8 alkyl;

[0014] R 2 is hydrogen or C1-C8 alkyl;

[0015] The C1-C8 alkyl mentioned above refers to a straight or branched alkyl group having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, pentyl, hexyl, heptyl, octyl, etc.; the halogen refers to fluorine, chlorine, bromine or iodine.

[0016] The B is a small molecule ligand of the CB protein in the E3 ubiquitin ligase complex, including amide compounds, phthalimide compounds, thalidomide or its derivatives, lenalidomide or its derivatives, pomalidomide or its derivatives. The general formula of some ligand structures is as follows:

[0017]

[0018] in:

[0019] G is selected from CH2, C=O, SO2, NH or N-alkyl;

[0020] X is selected from O or S;

[0021] Y is selected from NH, N-alkyl, N-aryl, N-heterocycle, N-aromatic heterocycle, N-cycloalkyl, O or S;

[0022] Z is selected from -alkyl, -cycloalkyl, halogen or hydrogen;

[0023] R 3 is selected from hydrogen, alkyl, hydroxy or -CH2-heterocycle;

[0024] W1, W2, W3, W4 are independently selected from nitrogen or carbon.

[0025] The halogen refers to fluorine, chlorine, bromine or iodine, preferably fluorine.

[0026] The L is a linking group selected from a nonlinear chain, an aliphatic chain, an aromatic chain, and a heteroaromatic ring structure chain, and is connected to A and B through a covalent bond. The structure of some linking groups is shown in the following general formula: n is an integer selected from 0 to 10,

[0027]

[0028] Furthermore, the compound is a compound as shown in Formula III:

[0029]

[0030] or a pharmaceutically acceptable salt thereof;

[0031] in:

[0032] (R) n Wherein R is halogen or C1-C5 alkyl, preferably halogen, and n is an integer selected from 0-2;

[0033] R 1 is hydrogen or C1-C5 alkyl, preferably hydrogen;

[0034] R 2 is hydrogen or C1-C5 alkyl, preferably hydrogen;

[0035] L is any one selected from the following structures: n1, n2, n3, n4 are independently selected from integers of 0-6,

[0036]

[0037] The halogen refers to fluorine, chlorine, bromine or iodine, preferably fluorine.

[0038] The C1-C5 alkyl group refers to a straight-chain or branched alkyl group having 1 to 5 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, and the like.

[0039] Furthermore, the compound is a compound as shown in Formula V:

[0040]

[0041] or a pharmaceutically acceptable salt thereof;

[0042] in:

[0043] L is any one selected from the following structures: n1, n2, n3, n4 are independently selected from integers of 0-6,

[0044]

[0045] Furthermore, the compound is one of the following compounds:

[0046]

[0047]

[0048] or a pharmaceutically acceptable salt thereof.

[0049] The present invention also provides the use of the compound and its pharmaceutically acceptable salt in the preparation of drugs for preventing and / or treating cancer. The chimeric molecule compound has the function of a PARP-1 inhibitor or a protein degrader. Furthermore, it has the dual functions of a PARP-1 inhibitor and a protein degrader, and can be used to prepare PARP-1 inhibitor or protein degrader drugs with the efficacy of preventing and treating cancer.

[0050] The present invention also provides the use of the compound or a pharmaceutically acceptable salt thereof in preventing and treating cancer, for example, the compound has cell proliferation inhibitory activity on breast cancer cell lines and / or ovarian cancer cell lines.

[0051] Furthermore, the compounds and pharmaceutically acceptable salts thereof of the present invention can be used alone as a sole anti-tumor drug, or can be used in combination with other active ingredients currently on the market for the treatment and prevention of cancer, etc.

[0052] Furthermore, the cancer may be, but is not limited to, multiple myeloma, gastric cancer, lung cancer, breast cancer, esophageal cancer, colon cancer, medulloblastoma, acute myeloid leukemia, chronic leukemia, prostate cancer, hepatoma, renal cell tumor, cervical cancer, skin cancer, ovarian cancer, colon cancer, glioma, thyroid cancer or pancreatic cancer.

[0053] The present invention also provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients. Such excipients include, but are not limited to, fillers, binders, disintegrants, release control agents, glidants, lubricants, coating agents, and the like that are used in the pharmaceutical field.

[0054] The present invention also provides a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof and other active ingredients. The other active ingredients are preferably selected from anticancer drugs, hormone drugs, interferon drugs, etc.

[0055] The invention also provides a preparation method for synthesizing the compound.

[0056] The compounds of the present invention also include their isomers and prodrugs, and may also be amorphous, anhydrous crystalline, hydrate crystalline, solvate crystalline or polymorphic forms of the compounds.

[0057] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined herein, they should be given the meaning that a person skilled in the art would give them based on the disclosure and context.

[0058] The term "substituted" refers to the replacement of a hydrogen atom in a molecule by another different atom or molecule.

[0059] The term "alkyl" refers to a straight-chain or branched alkyl group, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, sec-butyl, pentyl, hexyl, heptyl, octyl, etc.; the C1-C8 alkyl group refers to a straight-chain or branched alkyl group having 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, sec-butyl, pentyl, hexyl, heptyl, octyl, etc. The C1-C5 alkyl group refers to a straight-chain or branched alkyl group having 1 to 5 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, sec-butyl, pentyl, etc.

[0060] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0061] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically or physically compatible with the other ingredients that make up a pharmaceutical dosage form and physiologically compatible with the receptor.

[0062] The term "isomer" refers to enantiomers, diastereomers, racemates, cis-trans isomers, tautomers, geometric isomers, epimers and mixtures thereof of the above compounds.

[0063] The term "prodrug" refers to a precursor derivative of a compound that may have weak activity or even no activity, but after administration, can be converted into the corresponding biologically active form under physiological conditions (such as by metabolism, solvolysis or other means).

[0064] The terms "salts" and "pharmaceutically acceptable salts" refer to acidic and / or basic salts of the above compounds formed with inorganic and / or organic acids and bases, including zwitterionic salts (inner salts), and also quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final isolation and purification of the compound. They can also be obtained by mixing the above compounds, or their stereoisomers, with a suitable amount of acid or base (e.g., an equivalent amount). These salts may be precipitated in solution and collected by filtration, or recovered after evaporation of the solvent, or obtained by freeze-drying after reaction in an aqueous medium. The pharmaceutically acceptable salts of the present invention include addition salts formed with the following acids: for example, salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or salts with organic acids such as formic acid, acetic acid, propionic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, trifluoroacetic acid, maleic acid, fumaric acid, oxalic acid, benzoic acid, lactic acid, malic acid, tartaric acid, citric acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, p-toluenesulfonic acid, etc.; or salts with acidic amino acids such as aspartic acid, glutamic acid, etc.

[0065] Unless otherwise noted, conventional methods such as mass spectrometry, NMR, cell biology, and pharmacology within the skill of the art were employed. Unless otherwise noted, experimental materials, including culture media, cell lines, and experimental consumables, are commercially available or prepared using known methods. Unless specifically defined, nomenclature, laboratory procedures, and techniques related to analytical chemistry, synthetic chemistry, and medicinal and pharmaceutical chemistry described herein are known to those skilled in the art.

[0066] The present invention 1H NMR spectra were obtained using a Bruker instrument (400 MHz), and chemical shifts are expressed in ppm. Tetramethylsilane (0.00 ppm) was used as the internal standard. H NMR notation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of a doublet, dt = doublet of a triplet. Coupling constants, when provided, are in Hz.

[0067] The mass spectrum of the present invention is obtained by measuring with an LC / MS instrument, and the ionization mode can be ESI or APCI; the enzyme-linked immunosorbent assay instrument is Tecan Spark from Tecan Company.

[0068] Obviously, based on the above contents of the present invention, according to common technical knowledge and customary means in this field, without departing from the above basic technical ideas of the present invention, other various forms of modifications, replacements or changes can be made.

[0069] The following is a further detailed description of the above contents of the present invention through specific implementation methods in the form of embodiments, but this should not be understood as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above contents of the present invention fall within the scope of the present invention. DETAILED DESCRIPTION

[0070] The following examples and preparations are provided to further illustrate and illustrate the compounds of the present invention and their preparation methods. It should be understood that the scope of the following examples and preparations does not limit the scope of the present invention in any way.

[0071] Example 1. Preparation of Compound Cb-1

[0072]

[0073] F (30 g, 109.4 mmol) was added to a 1000 ml single-necked bottle, tert-butyl bromoacetate (42.7 g, 218.8 mmol) was added, potassium carbonate (30.2 g, 218.8 mmol) was added, 300 ml of DMF was added, and the mixture was reacted at 90°C overnight. After the reaction, the reaction solution was cooled, poured into 1500 ml of water, stirred, and filtered to obtain 40.1 g of G, which was used directly in the next step. 40.1 g of G was then added to 400 ml of formic acid and 100 ml of water and refluxed for 3 hours. After the reaction, the reaction solution was concentrated to 50 ml under reduced pressure, 500 ml of water was added, stirred, filtered, and dried. 28.8 g of Cb-1 was obtained, with a total yield of 78% for two steps, as a white solid powder. The characterization results are as follows: 1H NMR (300MHz, DMSO) δ13.48–13.00(m,1H),11.13(s,1H),7.80(dd,J=8.5,7.3Hz,1H),7.48(d,J=7.0Hz,1H),7.39(d, J=8.4Hz,1H),5.11(dd,J=12.8,5.3Hz,1H),5.00(s,2H),3.00–2.80(m,1H),2.69–2.53(m,2H),2.15–1.92(m,1H).

[0074] Example 2: Preparation of Compound Cb-2

[0075] (1) Synthesis of intermediate compound H

[0076]

[0077] T (2 g, 12.5 mmol) was added to a 250 ml round-bottom flask, followed by Cb-1 (4.15 g, 12.5 mmol), triethylamine (3.8 g, 37.5 mmol), HBTU (6.2 g, 18.75 mmol), and 100 ml of DMF. The mixture was stirred at room temperature for 2 hours. The reaction solution was poured into 500 ml of water and extracted with ethyl acetate three times with 200 ml each. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. H was purified by column chromatography using petroleum ether / ethyl acetate to obtain 4.3 g of H (yield 72.5%) as a white solid powder.

[0078] (2) Synthesis of Cb-2

[0079]

[0080] H (4.3 g, 9.1 mmol) was added to a 250 mL round-bottom flask, followed by 90 mL of dichloromethane and 11.3 mL of a 4N solution of hydrogen chloride in dioxane. The mixture was stirred at room temperature for 30 minutes, filtered, and air-dried to obtain 3.3 g of Cb-2 (89% yield) as a white solid powder. Characterization is as follows: 1H NMR(300MHz,dmso)δ11.12(s,1H),8.31(t,J=5.7Hz,1H),8.04(s,2H),7.81(dd,J=8.4,7.4Hz,1H),7.50(d,J=7.2Hz,1H),7.46(d,J=8.5H z,1H),5.12(dd,J=12.8,5.4Hz,1H),4.83(s,2H),3.40(dd,J=12.1,6.2Hz,2H),2.97–2.80(m,3H),2.67–2.53(m,1H),2.10–1.94(m,1H).

[0081] Example 3: Preparation of Compound C05

[0082] (1) Synthesis of intermediate compound A

[0083]

[0084] To a 50 mL reaction flask, add rucaparib (500 mg, 1.55 mmol) and potassium carbonate (428 mg, 3.09 mmol), add 10 mL of DMF, cool to 0°C, and slowly add ethyl bromoacetate (260 mg, 1.70 mmol) dropwise. The reaction is allowed to proceed at 25°C for 12 h. After completion, the reaction solution is poured into water and extracted with ethyl acetate. The combined organic phases are washed with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a pale yellow solid A in a 68.0% yield.

[0085] (2) Synthesis of intermediate compound B

[0086]

[0087] To a 50 mL reaction flask, intermediate A (200 mg, 0.50 mmol), 3 mL of methanol and 3 mL of tetrahydrofuran were added, followed by the addition of 2 mol·L -1 Aqueous LiOH solution (2.50 mL, 5.00 mmol) was added and reacted at room temperature overnight. After completion of the reaction, the reaction solution was dried by rotary evaporation, dissolved in 10 mL of water, and extracted with dichloromethane. The aqueous phase was neutralized with 1N hydrochloric acid to a pH of 5-6, filtered, and the filter cake was dried to obtain Compound B in a yield of 79.6%.

[0088] (3) Preparation of compound C05

[0089]

[0090] To a 10 mL reaction vial was added B (30 mg, 0.08 mmol), Cb-2 (33 mg, 0.08 mmol), and 2 mL of DMF. HOBt (12 mg, 0.09 mmol), EDCI (18 mg, 0.09 mmol), and DIEA (50 mg, 0.40 mmol) were also added. The reaction was allowed to proceed at 25°C for 12 h. The reaction solution was poured into water, and a solid precipitated. This was filtered and dried to yield 26.90 mg of an off-white solid, with a total yield of 22.5%. The structure of C05 is as follows:

[0091]

[0092] The characterization results are as follows: 1 H NMR (500MHz, DMSO-d6) δ11.65(s,1H),11.11(s,1H),8.25(q,J=6.4,5.9Hz,1H),8.04(q,J=5.6Hz,1H ),7.95(t,J=5.5Hz,1H),7.75(t,J=7.9Hz,1H),7.59–7.49(m,4H),7.48–7.28(m,4H),5.10(dd,J=12 .8,5.5Hz,1H),4.75(s,2H),3.59(d,J=9.0Hz,4H),3.26(dt,J=10.5,5.7Hz,3H),2.99–2.83(m,4H), 2.63–2.52(m,2H),2.16(s,3H),2.00(dt,J=13.5,3.5Hz,1H),1.81–1.66(m,2H).MS:737.79(M+H+).

[0093] Example 4: Preparation of Compound C06

[0094]

[0095] To a 10 mL reaction flask, rucaparib (40 mg, 0.12 mmol), Cb-1 (49 mg, 0.15 mmol), and 2 mL of DMF were added. HOBt (17 mg, 0.14 mmol), EDCI (24 mg, 0.14 mmol), and DIEA (68 mg, 0.52 mmol) were then added and allowed to react at 25°C for 12 h. The reaction solution was poured into water, and the solid washed out, filtered, and dried to yield 49.76 mg of an off-white solid, C06, in a 65% yield. Characterization is as follows: 1H NMR (500MHz, DMSO-d6) δ11.68(s,1H),11.11(d,J=2.8Hz,1H),8.26(t,J=5.7Hz,1H ),7.95(s,1H),7.80(dd,J=9.6,6.3Hz,1H),7.68–7.55(m,2H),7.50–7.38(m,4H), 7.34–7.31(m,1H),5.11(dd,J=12.6,5.4Hz,1H),4.76–4.35(m,2H),3.04(s,3H),2 .89(d,J=2.3Hz,4H),2.73(d,J=2.2Hz,4H),2.28–1.85(m,2H).MS: 638.12(M+H+).

[0096] Example 5: Preparation of Compound C07

[0097] (1) Synthesis of intermediate compound Cb-3

[0098] The synthesis of Cb-3 refers to Example 2, using T-1 instead of T.

[0099]

[0100] Get Cb-3,

[0101]

[0102] Characterization is as follows: 1 H NMR(300MHz,dmso)δ11.12(s,1H),8.23(t,J=5.8Hz,1H),7.96(s,3H),7.82(dd,1H),7.50(d,J=7.2Hz,1H),7.40(d,J=8.5Hz,1H),5.12(dd,J=12 .8,5.5Hz,1H),4.81(s,2H),3.29–3.12(m,2H),3.00–2.82(m,1H),2.83 –2.71(m,2H),2.65–2.53(m,2H),2.13–1.97(m,1H),1.86–1.65(m,2H).

[0103] (2) Synthesis of compound C07

[0104] The specific preparation method is as shown in Example 3, except that Cb-2 in Example 3 is replaced by Cb-3. 28.60 mg of an off-white solid C07 is obtained with a total yield of 24.5%. The structure of C07 is as follows:

[0105]

[0106] The characterization results are as follows: 1 H NMR (500MHz, DMSO-d6) δ11.67(s,1H),11.11(s,1H),8.25(t,J=5.8Hz,1H),8.03(t,J=5.9Hz,1H),7.92(t,J=6.1Hz,1H),7. 80(t,J=7.9Hz,1H),7.62–7.52(m,4H),7.48(d,J=7.4Hz,1H),7.41(dd,J=12.5,9.0Hz,2H),7.34–7.26(m,1H),5.11(dd,J=1 2.7,5.5Hz,1H),4.79(s,2H),3.62(s,2H),3.15(dq,J=18.7,6.8Hz,6H),3.11–3.00(m,2H),2.97(s,2H),2.88(ddd,J=18.0 ,13.6,5.4Hz,1H),2.62–2.55(m,2H),2.21(s,3H),2.01(dd,J=12.4,6.3Hz,1H),1.59(p,J=6.9Hz,2H).MS: 751.97(M+H+).

[0107] Example 6: Preparation of Compound C08

[0108] (1) Synthesis of intermediate compound Cb-4

[0109] The synthesis of Cb-4 refers to Example 2, using T-2 instead of T.

[0110]

[0111] Get Cb-4,

[0112]

[0113] Characterization is as follows: 1 H NMR(300MHz,dmso)δ11.12(s,1H),8.14(t,J=7.5,4.0Hz,1H),8.01(s,2H),7. 83(dd,J=8.4,7.4Hz,1H),7.50(d,J=7.2Hz,1H),7.41(d,J=8.5Hz,1H),5.12( dd,J=12.9,5.4Hz,1H),4.79(s,2H),3.16(q,J=6.1Hz,2H),2.99–2.84(m,1H) ,2.83–2.68(m,2H),2.67–2.53(m,2H),2.17–1.96(m,1H),1.76–1.32(m,4H).

[0114] (2) Synthesis of compound C08

[0115] C08 was prepared. The specific preparation method was as described in Example 3. Cb-2 in Example 3 was replaced with Cb-4. 25.80 mg of an off-white solid C08 was obtained in accordance with Example 3. The total yield was 22.8%. The structure of C08 is as follows:

[0116]

[0117] The characterization results are as follows: 1 H NMR(500MHz,DMSO-d6)δ11.67(s,1H),11.12(s,1H),8.25(d,J=5.9Hz,1H),8.07– 7.92(m,1H),7.81(dt,J=32.2,7.0Hz,2H),7.56(dd,J=34.3,7.7Hz,4H),7.39(ddd , J=53.3,21.8,8.2Hz,4H),5.23–5.10(m,1H),4.74(d,J=11.3Hz,2H),3.61(s,2H) ,3.27–2.54(m,12H),2.20(s,3H),1.44(s,4H),1.27–0.61(m,2H).MS:766.02(M+H + ).。

[0118] Example 7: Preparation of Compound C09

[0119] (1) Synthesis of intermediate compound Cb-5

[0120] The synthesis of Cb-5 refers to Example 2, using T-3 instead of T.

[0121]

[0122] Get Cb-5,

[0123]

[0124] Characterization is as follows: 1H NMR (300MHz, Methanol-d4) δ7.82 (dd, J=8.5, 7.3Hz, 1H), 7.54 (dd, J=7.4, 0.6Hz, 1H), 7.44 (dd, J=8.5, 0.6Hz, 1H), 5.14 (dd, J=12.5 ,5.4Hz,1H),4.78(s,2H),3.72–3.58(m,12H),3.51(t,J=5.2Hz,2H),3.12(t,J=5.0Hz,2H),2.99–2.62(m,3H),2.22–2.08(m,1H).

[0125] (2) Synthesis of compound C09

[0126] C09 was prepared. The specific preparation method was as described in Example 3. Cb-2 in Example 3 was replaced with Cb-5. 28.40 mg of C09 as an off-white solid was obtained in accordance with Example 3. The total yield was 25.8%. The structure of C09 is as follows:

[0127]

[0128] The characterization results are as follows: 1 H NMR(500MHz,DMSO-d6)δ11.69(s,1H),11.12(s,1H),8.26(s,1H),8.00(s,1H),7.81(s,2H),7.63–7.19(m,8H), 5.11(s,1H),4.77(s,2H),3.80–3.18(m,21H),2.97(t,J=39.3Hz,5H),2.11(s,3H),1.76(s,2H).MS: 869.97(M+H + ).。

[0129] Example 8: Preparation of Cb-6

[0130] (1) Synthesis of intermediate compound J

[0131]

[0132] T (4 g, 25 mmol) was added to a 250 ml round-bottom flask, followed by I (6.9 g, 25 mmol) and diisopropylethylamine (6.5 g, 50 mmol). 80 ml of DMF was added, and the mixture was stirred at 90°C for 2 hours. The reaction solution was poured into 500 ml of water and extracted with ethyl acetate three times with 200 ml each. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 4.3 g of J was obtained by column chromatography using petroleum ether / ethyl acetate, with a yield of 41%, as a dark green solid powder.

[0133] (2) Synthesis of intermediate compound Cb-6

[0134]

[0135] J (4.3 g, 10.3 mmol) was added to a 250 ml round-bottom flask, and 90 ml of dichloromethane was added, followed by 13 ml of a 4N hydrogen chloride solution in dioxane. The mixture was stirred at room temperature for 30 minutes, filtered, and air-dried to obtain 3.4 g of Cb-6 (yield 93%) as a yellow solid powder.

[0136] Example 9: Preparation of Compound C10

[0137] C10 was prepared by referring to Example 3 for the specific preparation method, except that Cb-2 in Example 3 was replaced with Cb-6. 26.80 mg of C10 as an off-white solid was obtained in accordance with Example 3 with a total yield of 24.6%. The structure of C10 is as follows:

[0138]

[0139] The characterization results are as follows: 1 H NMR (500MHz, DMSO-d6) δ11.66(s,1H),11.09(s,1H),8.26(t,J=5.8Hz,1H),8.10(t,J=5.8Hz,1H),7.57(dt,J=8.9,3.7Hz,3H),7.5 0(d,J=8.2Hz,2H),7.42(dd,J=11.0,2.4Hz,1H),7.32(dd,J=9.0,2.5Hz,1H),7.22(d,J=8.6Hz,1H),7.01(d,J=7.0Hz,1H),6.74(t ,J=6.0Hz,1H),5.03(dd,J=12.8,5.4Hz,1H),3.59(s,2H),3.42(s,2H),3.03(q,J=4.8,3.9Hz,2H),2.97(d,J=8.6Hz,3H),2.89(s, 1H),2.87–2.81(m,1H),2.73(s,1H),2.55(t,J=3.4Hz,1H),2.42(q,J=7.1Hz,2H),2.18(s,3H),2.00–1.93(m,1H).MS: 679.95(M+H + ).。

[0140] Example 10: Preparation of Compound C11

[0141] (1) Synthesis of intermediate compound Cb-7

[0142] The synthesis of Cb-7 was carried out according to Example 8, except that T-1 was used to replace T.

[0143]

[0144] Get Cb-7,

[0145]

[0146] Characterization is as follows: 1 H NMR(300MHz,dmso)δ11.00(s,1H),8.01–7.75(m,2H),7.50(dd,J=8.5,7.1Hz,1H),7.06(d,J=8.5Hz,1H),6.94(d,J=6.9Hz,1H),6.66(t,J=6.5H z,1H),4.95(dd,J=12.9,5.4Hz,1H),3.39–3.27(m,2H),2.76(dd,J=12. 8, 4.3Hz, 3H), 2.55–2.41 (m, 2H), 2.01–1.86 (m, 1H), 1.83–1.65 (m, 2H).

[0147] (2) Synthesis of compound C11

[0148] C11 was prepared by referring to Example 3. Cb-2 in Example 3 was replaced with Cb-7. 24.30 mg of an off-white solid C11 was obtained in accordance with Example 3 with a total yield of 22.4%. The structure of C11 is as follows:

[0149]

[0150] The characterization results are as follows: 1H NMR (500MHz, DMSO-d6) δ11.67(s,1H),11.09(s,1H),8.25(t,J=5.6Hz,1H),8.02(t,J=6.1Hz,1H),7.59(d,J=8.0Hz,2H),7.57–7. 50(m,3H),7.42(dd,J=11.0,2.5Hz,1H),7.32(dd,J=9.1,2.5Hz,1H),7.06(d,J=8.6Hz,1H),7.00(d,J=7.0Hz,1H),6.76(t,J=6.2 Hz,1H),5.05(dd,J=12.7,5.4Hz,1H),3.62(s,2H),3.38(s,1H),3.21(q,J=6.4Hz,2H),3.06–3.02(m,2H),2.99(s,2H),2.89(s,2 H),2.73(s,1H),2.61–2.52(m,2H),2.22(s,3H),2.05–2.00(m,1H),1.71(p,J=6.7Hz,2H),0.95(d,J=6.6Hz,3H).MS: 693.82(M+H + ).。

[0151] Example 11: Preparation of Compound C12

[0152] (1) Synthesis of intermediate compound Cb-8

[0153] The synthesis of Cb-8 refers to Example 8, using T-2 instead of T.

[0154]

[0155] Get Cb-8,

[0156]

[0157] Characterization is as follows: 1 H NMR(300MHz,dmso)δ11.01(s,1H),7.78(s,2H),7.49(dd,J=8.5,7.1Hz,1H),7.08–6.94(m,1H),6.94(d,J=6.9Hz,1H),6.51(t,J=6.1Hz,1H),4.9 5(dd,J=12.8,5.4Hz,1H),3.35–3.25(m,2H),2.88–2.76(m,1H),2.73(d, J=7.4Hz,2H),2.55–2.39(m,2H),2.06–1.83(m,1H),1.62–1.45(m,4H).

[0158] (2) Synthesis of compound C12

[0159] C12 was prepared by referring to Example 3. Cb-2 in Example 3 was replaced with Cb-8. 28.20 mg of an off-white solid C12 was obtained in accordance with Example 3 with a total yield of 26.5%. The structure of C12 is as follows:

[0160]

[0161] The characterization results are as follows: 1 H NMR (500MHz, DMSO-d6) δ11.67(s,1H),11.03(s,1H),8.25(t,J=5.8Hz,1H),7.89(t,J=6.1Hz,1H),7.59(d,J=8.2Hz,2H),7.52(d,J=7. 8Hz,3H),7.42(dd,J=11.0,2.4Hz,1H),7.32(dd,J=9.1,2.5Hz,1H),7.07(d,J=8.6Hz,1H),6.99(d,J=7.0Hz,1H),6.56(t,J=6.0Hz,1H) ,5.03(dd,J=12.7,5.5Hz,1H),3.61(s,2H),3.38(s,1H),3.29(d,J=6.6Hz,3H),3.16(q,J=6.4Hz,2H),3.06–3.01(m,2H),2.97(s,2H), 2.89(s,1H),2.87–2.82(m,1H),2.56(ddd,J=16.8,4.4,2.3Hz,1H),2.19(s,3H),2.03–1.96(m,1H),1.59–1.49(m,4H).MS: 707.80(M+H + ).。

[0162] Example 12: Preparation of Compound C13

[0163] (1) Synthesis of intermediate compound Cb-9

[0164] The synthesis of Cb-9 refers to Example 8, using T-4 instead of T.

[0165]

[0166] Get Cb-9,

[0167]

[0168] Characterization is as follows: 1H NMR(300MHz,dmso)δ11.00(s,1H),7.85(s,2H),7.49(dd,J=8.6,7.1Hz,1H),7 .00(d,J=8.5Hz,1H),6.92(d,J=7.0Hz,1H),6.59–6.28(m,1H),4.95(dd,J=12. 9,5.4Hz,1H),3.20(t,J=6.7Hz,2H),2.76(d,J=13.6Hz,1H),2.69–2.57(m,2H) ,2.56–2.41(m,2H),2.05–1.81(m,1H),1.60–1.36(m,4H),1.32–1.14(m,4H).

[0169] (2) Synthesis of compound C13

[0170] C13 was prepared by referring to Example 3 for the specific preparation method. Cb-2 in Example 3 was replaced with Cb-9. 25.30 mg of C13 as an off-white solid was obtained in accordance with Example 3 with a total yield of 25.5%. The structure of C13 is as follows:

[0171]

[0172] The characterization results are as follows: 1 H NMR (500MHz, DMSO-d6) δ11.68(s,1H),10.90(s,1H),8.25(t,J=5.8Hz,1H),7.81(t,J=6.0Hz,1H),7.59(d,J=8.0Hz,2H),7.56–7.50(m,3H),7.42(d d,J=11.0,2.4Hz,1H),7.31(dd,J=9.0,2.5Hz,1H),7.03(d,J=8.6Hz,1H),6.99(d,J=7.1Hz,1H),6.50(t,J=5.9Hz,1H),5.04(dd,J=12.8,5.5Hz,1H ),3.61(s,2H),3.24(t,J=6.7Hz,2H),3.11(q,J=6.7Hz,2H),3.04(q,J=6 .3,5.6Hz,2H),2.96(s,2H),2.90–2.87(m,1H),2.61–2.55(m,1H),2.47–2 .44(m,1H),2.20(s,3H),2.04–1.97(m,1H),1.54(t,J=7.3Hz,2H),1.44( q,J=7.1Hz,2H),1.37–1.23(m,5H),0.94(d,J=6.5Hz,1H).MS:736.03(M+H +).。

[0173] Example 13: Preparation of Compound C14

[0174] (1) Synthesis of intermediate compound Cb-10

[0175] The synthesis of Cb-10 refers to Example 8, using T-5 instead of T.

[0176]

[0177] Get Cb-10,

[0178]

[0179] Characterization is as follows: 1 H NMR(300MHz,dmso)δ11.10(s,1H),7.95(s,2H),7.58(dd,J=8.5,7.1Hz,1H), 7.09(d,J=8.6Hz,1H),7.02(d,J=7.0Hz,1H),6.62–6.45(m,1H),5.05(dd,J=1 2.8,5.4Hz,1H),3.29(d,J=6.0Hz,2H),2.98–2.81(m,1H),2.79–2.67(m,2H), 2.67–2.49(m,2H),2.14–1.94(m,1H),1.66–1.43(m,4H),1.42–1.19(m,8H).

[0180] (2) Synthesis of compound C14

[0181] C14 was prepared by referring to Example 3 for the specific preparation method. Cb-2 in Example 3 was replaced with Cb-10. 23.80 mg of C14 as an off-white solid was obtained in accordance with Example 3 with a total yield of 24.1%. The structure of C14 is as follows:

[0182]

[0183] The characterization results are as follows: 1H NMR (500MHz, DMSO-d6) δ11.67(s,1H),11.09(s,1H),8.25(t,J=5.8Hz,1H),7.80(t,J=5.9Hz,1H),7.60(d,J=8.2Hz,2H),7.53(t,J=7.1H z,3H),7.42(dd,J=10.9,2.4Hz,1H),7.31(dd,J=9.1,2.4Hz,1H),7.02(dd,J=17.5,7.8Hz,2H),6.48(t,J=5.9Hz,1H),5.04(dd,J=12.8,5 .4Hz,1H),3.61(s,2H),3.39(s,1H),3.23(q,J=6.7Hz,3H),3.10(q,J=6.6Hz,2H),3.04(q,J=7.4,6.3Hz,2H),2.96(s,2H),2.89(s,1H),2 .62–2.55(m,1H),2.20(s,3H),2.02(ddd,J=12.7,5.4,3.0Hz,1H),1.55–1.49(m,2H),1.41(d,J=6.9Hz,2H),1.25(s,9H).MS:764.07(M+H + ).。

[0184] Example 14: Preparation of Compound C15

[0185] (1) Synthesis of intermediate compound Cb-11

[0186] The synthesis of Cb-11 refers to Example 2, using T-5 instead of T.

[0187]

[0188] Get Cb-11,

[0189]

[0190] Characterization is as follows: 1H NMR(300MHz,dmso)δ11.12(s,1H),7.96(t,J=5.6Hz,1H),7.87–7.77(m,1H),7.7 2(s,2H),7.51(d,J=7.2Hz,1H),7.40(d,J=8.5Hz,1H),5.12(dd,J=12.8,5.2Hz,1 H),4.77(s,2H),3.14(dd,J=12.6,6.4Hz,2H),3.00–2.82(m,1H),2.83–2.67(m,2 H),2.65–2.52(m,2H),2.17–1.94(m,1H),1.62–1.36(m,4H),1.36–1.13(m,8H).

[0191] (2) Synthesis of compound C15

[0192] C15 was prepared by referring to Example 3 for the specific preparation method. Cb-2 in Example 3 was replaced with Cb-11. 26.40 mg of C15 as an off-white solid was obtained in accordance with Example 3 with a total yield of 26.1%. The structure of C15 is as follows:

[0193]

[0194] The characterization results are as follows: 1 H NMR (500MHz, DMSO-d6) δ11.67(s,1H),11.12(s,1H),8.25(t,J=5.8Hz,1H),7.90(t,J=5.9Hz,1H),7.81(d,J=7.4Hz,2H),7.60(d,J=7.9 Hz,2H),7.53(d,J=8.0Hz,2H),7.49(d,J=7.2Hz,1H),7.42(dd,J=10.9,2.5Hz,1H),7.38(d,J=8.5Hz,1H),7.31(dd,J=9.2,2.5Hz,1H),5 .11(dd,J=12.8,5.5Hz,1H),4.75(s,2H),3.61(s,2H),3.38(d,J=6.8Hz,2H),3.11(q,J=6.8Hz,4H),3.06–3.03(m,2H),2.96(s,1H),2. 89(s,1H),2.73(s,1H),2.59(d,J=19.9Hz,2H),2.20(s,3H),2.03(d,J=12.6Hz,1H),1.40(d,J=7.5Hz,3H),1.23(s,9H).MS: 822.03(M+H + ).。

[0195] Example 15: Biological assay of the inhibitory effect of the compounds of the present invention on the proliferation of breast cancer cells MDA-MB-231

[0196] (1) Experimental principle: Cell Counting Kit-8, also known as CCK-8 kit, is a rapid and highly sensitive detection kit based on WST-8 and widely used in cell proliferation and cytotoxicity.

[0197] The CCK-8 reagent contains WST-8 (chemical name: 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid benzene)-2H-tetrazolium monosodium salt). This is reduced by dehydrogenases in the mitochondria under the action of electron carriers to produce orange-yellow formazan. The more and faster the cell proliferation, the darker the color; the greater the cytotoxicity, the lighter the color. For the same cells, the color intensity is linearly correlated with cell number. The number of viable cells is determined by the measured absorbance (OD value). A higher OD value indicates greater cell viability. For drug toxicity testing, a higher OD value indicates less toxicity.

[0198] (2) Reagents and cells:

[0199] Reagents: DMEM medium: CellMax CGN101.5, supplemented with 10% FBS (Gemini 900-108); CCK-8 kit: Bimake 25 mL.

[0200] Test cells: breast cancer cells MDA-MB-231 (purchased from: Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences)

[0201] (3) Experimental steps:

[0202] 1) Collect logarithmic phase cells, adjust the cell suspension concentration with DMEM medium, and aliquot into 96-well plates, 100 μL per well, 3,000-10,000 cells / well;

[0203] 2) Place in a 37°C, 5% CO2 incubator to allow cells to adhere for 24 hours;

[0204] 3) Aspirate the culture medium, wash three times with PBS, dissolve the test compound in DMEM medium, add 200 μL of the culture medium containing the test compound to each well (concentration 2 μM, triplicate wells), and continue culturing for 48 h;

[0205] 4) Add 10 μL of CCK-8 reagent to each well and continue incubation for 4 h;

[0206] 5) Measure the absorbance of each well at 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader;

[0207] 6) Set up blank wells (DMEM medium, CCK-8 reagent) and control wells (cells, DMEM medium, CCK-8 reagent) at the same time, with three replicates per group;

[0208] 7) Calculate the inhibition rate = [(OD 对照 -OD 空白 )-(OD 给药 -OD 空白 )] / (OD 对照 -OD 空白 )*100%.

[0209] The comparison table of the defined inhibition rate range and inhibition degree is as follows:

[0210] Inhibition rate (%) Degree of inhibition 0-5％ + 5-10％ ++ 10-15％ +++ 15-20％ ++++ More than 20% +++++ No data obtained /

[0211] Note: Inhibition rate 5-10% refers to the inhibition rate range of more than 5% and less than or equal to 10%, and so on.

[0212] Table 1. Activity data of the compounds of the present invention on the proliferation inhibition of breast cancer cells MDA-MB-231

[0213]

[0214]

[0215]

[0216] The above results demonstrate that compounds C06, C09, C10, C13, C14, and C15 disclosed herein exhibit stronger cell proliferation inhibitory activity against MDA-MB-231 breast cancer cells than rucaparib, demonstrating that the chimeric molecules possess superior anti-tumor efficacy compared to rucaparib, potentially reducing drug dosage and alleviating toxic side effects. Furthermore, compound C05 exhibited comparable cytostatic activity against MDA-MB-231 breast cancer cells to that of rucaparib.

[0217] Example 16: Biological assay of the inhibitory effect of the compounds of the present invention on ovarian cancer cell A2780 cell proliferation

[0218] (1) Experimental principle: Same as Example 15

[0219] (2) Reagents and cells:

[0220] Reagents: DMEM medium: CellMax CGN101.5, supplemented with 10% FBS (Gemini 900-108); CCK-8 kit: Bimake 25 mL.

[0221] Test cells: ovarian cancer cell A2780 cells (purchased from: Cell Resource Center, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences).

[0222] (3) Experimental procedures: Same as Example 15.

[0223] Table 2. Activity data of the compounds of the present invention on the inhibition of proliferation of ovarian cancer cell A2780

[0224]

[0225]

[0226]

[0227] These results demonstrate that compounds C05, C07, C13, and C14 disclosed herein exhibit stronger cell proliferation inhibitory activity against ovarian cancer cell lines A2780 than rucaparib, demonstrating that the chimeric molecules C05, C07, C13, and C14 possess superior anti-tumor efficacy compared to the PARP-1 inhibitor rucaparib, potentially reducing drug dosage and alleviating toxic side effects. Furthermore, compounds C06, C08, C09, C10, C11, C12, and C15 exhibit comparable cytostatic activity against ovarian cancer cells A2780 as rucaparib.

[0228] In summary, the present invention provides a chimeric molecule, which is composed of a target protein-targeting small molecule compound unit, an E3 ubiquitin ligase binding unit and a linker group. It has cancer cell proliferation inhibitory activity and can be used as a PARP-1 protein degradation and / or inhibition drug for the treatment of cancer.

Claims

1. Compounds or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, characterized in that The pharmaceutically acceptable salt is an acid addition salt, and the acid used for the addition salt is selected from: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, naphthalene disulfonic acid, acetic acid, propionic acid, pyruvic acid, lactic acid, trifluoroacetic acid, maleic acid, citric acid, fumaric acid, oxalic acid, tartaric acid, benzoic acid, and succinic acid.

3. Use of the compound according to claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and / or treating breast cancer and / or ovarian cancer.

4. A pharmaceutical composition, characterized in that: The invention comprises the compound according to claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

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