PROTAC molecules based on AZD5305 derivation as well as preparation method and application of PROTAC molecules

Abstract

The invention discloses a PROTAC molecule derived based on AZD5305 as well as a preparation method and application of the PROTAC molecule. AZD5305 and a CRBN ligand are connected through a linker, a series of PROTAC molecules derived from AZD5305 are provided, and the compound can be used as a degradation agent for targeting PARP1, can play a role in efficiently degrading PARP1 protein, and is used for treating cancers such as triple negative breast cancer and the like. Therefore, the compound provided by the invention can be used for preparing a PARP1 degradation agent and can be used for preparing drugs for treating cancers.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a class of PROTAC molecules derived from AZD5305 and a preparation method and use thereof. Background Art

[0002] Poly (ADP-ribose) polymerase 1 (PARP1) is a nuclear protein that is mainly responsible for poly ADP-ribosylation modification in organisms. It can recruit many DNA repair factors and promote DNA repair. PARP1 mainly mediates single-strand repair of DNA in the body. If the loss of single-strand DNA is not repaired in time, double-strand DNA damage will occur. Normal cells can repair double-strand DNA damage through HR recombination in the body to ensure the stability of the genome, but some tumor cells lack HR protein. Inhibiting PARP1 protein will induce cells to produce more toxic double-strand DNA damage, thereby killing tumor cells. This mechanism is called synthetic lethality.

[0003] PARP1 inhibitors (PARPi) have made great progress since 2010, but inhibitors still have some defects, such as the clinical toxicity of PARPi, which is caused by the trapping mechanism of PARPi (Yves Pommier et al. Laying a trap to kill cancer cells: PARP inhibitors and their mechanisms of action. Sci. Transl. Med. 8, 362ps17-362ps17 (2016).), which limits the clinical application of PARPi in other diseases.

[0004] A variety of inhibitors have been developed based on PARP1, but due to the potential toxicity of the capture characteristics of PARP1 inhibitors, their clinical use is limited, and drug resistance often occurs in the use of clinical inhibitors. PROTAC molecules have the characteristics of catalytic degradation and can degrade proteins at low doses, which can overcome the drug resistance of PARP1 inhibitors used clinically. In addition, studies have shown that PARP1 degraders can inhibit the skeleton function and catalytic function of PARP1, providing potential applications for the study of the mechanism of PARP1 in diseases.

[0005] AZD5305 is a new generation of PARP inhibitor from AstraZeneca, and its structural formula is as follows:

[0006]

[0007] AZD5305 is a second-generation PARP1 inhibitor. Due to its high selectivity for PARP1, it can avoid the side effects caused by the inhibition of PARP2. Previous PARP1 degraders were based on first-generation inhibitors as targeted warheads. Their poor selectivity for PARP1 / PARP2 may bring potential risks in clinical use. PROTAC designed based on second-generation PARP1 inhibitors may provide a solution. Summary of the invention

[0008] Objective of the invention: One of the objectives of the present invention is to provide a PROTAC molecule derived from AZD5305 or a pharmaceutically acceptable salt thereof according to general formula I:

[0009]

[0010] in,

[0011] L is a linker selected from a C5-C12 aliphatic chain, a saturated heterocycle or an alkyl chain containing 2-4 N atoms;

[0012] D is selected from:

[0013]

[0014] A preferred embodiment of the present invention is:

[0015] L is selected from:

[0016]

[0017] In some preferred embodiments, the pharmaceutically acceptable salts include, but are not limited to, acid addition salts formed between the compound of formula I and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid; and also include acid salts formed between the compound of formula I and inorganic bases.

[0018] The compounds of the general formula I of the present invention are preferably the following compounds:

[0019]

[0020]

[0021] The above-mentioned compounds of general formula I of the present invention may also exist in the form of their salts, which are converted in vivo into compounds of general formula I. For example, within the scope of the present invention, the compounds of the present invention are converted into pharmaceutically acceptable salt forms according to processes known in the art, and they are used in the form of salts.

[0022] Another object of the present invention is to provide a method for preparing a compound of formula I, comprising the following steps:

[0023]

[0024] (1) Compound A undergoes a nucleophilic reaction with an aliphatic hydrocarbon chain, a heteroaromatic ring structure compound, or an alkyl chain compound containing 2 to 4 N atoms, or undergoes a condensation reaction in the presence of a condensation agent to obtain Compound B;

[0025] (2) Compound B is subjected to acid hydrolysis to remove the Boc protecting group, and then undergoes a condensation reaction with compound C in the presence of an amide condensation agent to generate a compound of formula I.

[0026] wherein X is selected from H, O, and L and D are as defined in Formula I.

[0027] Compound C was prepared according to the method in CN 117177972A.

[0028] The compounds of the general formula I of the present invention can be prepared by the above or similar preparation methods, and the corresponding starting materials can be selected according to the different substituents and the different positions of the substituents. It should be appreciated by those skilled in the art that the above routes are helpful for understanding the present invention, but do not limit the content of the present invention. Unless otherwise specified, the variables are defined as mentioned in the general formula I.

[0029] Another object of the present invention is to provide a pharmaceutical composition, which comprises a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0030] The dosage form of the pharmaceutical composition of the present invention can be tablets, capsules, powders, syrups, liquids, suspensions, lyophilized powders or injections.

[0031] The pharmaceutical composition of the present invention can be administered in various known ways, such as orally, parenterally, by inhalation spray or via an implanted reservoir. The pharmaceutical composition of the present invention can be administered alone or in combination with other drugs. Oral compositions can be in any orally acceptable dosage form, including but not limited to tablets, capsules, powders, syrups, liquids and suspensions. Commonly used pharmaceutically acceptable carriers include stabilizers, diluents, surfactants, lubricants, antioxidants, adhesives, colorants, fillers, emulsifiers, etc.

[0032] The actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient. The dosage level selected depends on a variety of factors, including the activity of the specific compound of the present invention or its salt used, the route of administration, the time of administration, the excretion rate of the specific composition used, the duration of treatment, other drugs, compounds and / or materials used in combination with the specific composition used, the age, sex, weight, general health and previous medical history of the patient being treated, and similar factors well known in the medical field.

[0033] Another object of the present invention is to provide use of the compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a PARP1 degrader.

[0034] Another object of the present invention is to provide use of the compound of formula I or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating cancer.

[0035] Such cancers include, but are not limited to, triple-negative breast cancer.

[0036] Beneficial effects:

[0037] (1) Based on PROTAC technology, the present invention connects AZD5305 and CRBN ligand through a linker, providing a series of PROTAC molecules derived from AZD5305. Such compounds can be used as PARP1-targeting degraders, and can effectively degrade PARP1 protein for the treatment of triple-negative breast cancer and other cancers.

[0038] (2) The compounds of the present invention can significantly degrade the expression level of PARP1 protein under MDA-MB-231, among which compounds I-12 and I-13 can degrade >85% of PARP1 protein at the nanomolar level, and can be used as chemical knockout tools for PARP1, as well as candidate drugs for anti-breast cancer. DETAILED DESCRIPTION

[0039] The preparation method of the compound of general formula I of the present invention is described below in conjunction with specific examples, but these specific methods do not constitute any limitation to the present invention. The compound of the present invention can also be conveniently prepared by optionally combining various synthetic methods described in this specification or known in the art, and such a combination can be easily carried out by a technician in the field to which the present invention belongs.

[0040] The starting materials, reaction reagents, etc. used in the specific embodiments of the present invention are all commercially available. The present invention can be prepared in the form of salt by using a salt-forming method commonly used in the art, for example, dissolving the compound in hydrochloric acid ethanol at room temperature to react to generate a hydrochloride; or adding benzenesulfonic acid to react to generate a benzenesulfonate.

[0041] Compound C was prepared according to the method in CN 117177972A.

[0042] Example 1: Preparation of N-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)amino)-5-oxopentyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-1)

[0043]

[0044] 261 mg (1.2 mmol) of Boc-5-aminovaleric acid and 495 mg (1.3 mmol) of HATU were dissolved in 5 ml of DMF, 523 μl (3 mmol) of DIPEA were added, and after stirring for 10 min, 260 mg (1 mmol) of lenalidomide (CAS No. 191732-72-6) was added, and the mixture was stirred at room temperature overnight. The reaction solution was extracted with EA and water, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-1 (413 mg, yield 90%) was obtained by column chromatography.

[0045] Compound A-1 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1, V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, and stirred until dissolved. 0.2 mmol HATU and 0.15 mmol compound C were added, and stirred at room temperature overnight. 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH. Compound I-1 (46 mg, yield 41.7%) was obtained by silica gel column chromatography.

[0046] 1 H NMR(300MHz,DMSO)δ(ppm)11.88(s,1H),11.04(s,1H),9.79(s,1H),8.45-8.36(m,2 H),8.25(d,J=2.8Hz,1H),7.86-7.77(m,2H),7.75(s,1H),7.63(d,J=1.9Hz,1H),7. 53-7.43(m,2H),7.38(dd,J=8.9,2.9Hz,1H),5.14(dd,J=13.3,5.1Hz,1H),4.44-4. 27(m,2H),3.64(s,2H),3.32(s,3H),3.25(d,J=6.5Hz,2H),2.98-2.84(m,1H),2.65

[0047] -2.52(m,6H),2.34(t,J=7.6Hz,3H),2.01(d,J=12.5Hz,1H),1.54(dt,J=27.2,7.0Hz,4H),1.30(d,J=3.2Hz,4H),1.17(t,J=7.4Hz,3H).

[0048] Example 2: Preparation of N-(6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)amino)-6-oxohexyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-2)

[0049]

[0050] 278 mg (1.2 mmol) of Boc-6-aminohexanoic acid and 495 mg (1.3 mmol) of HATU were dissolved in 5 ml of DMF, 523 μl (3 mmol) of DIPEA were added, and after stirring for 10 min, 260 mg (1 mmol) of lenalidomide was added, and the mixture was stirred at room temperature overnight. The reaction solution was extracted with EA and water, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-2 (412 mg, yield 87%) was obtained by column chromatography.

[0051] Compound A-2 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1, V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, and stirred until dissolved. 0.2 mmol HATU and 0.15 mmol compound C were added, and stirred at room temperature overnight. 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH. Compound I-2 (52 mg, yield 46.3%) was obtained by silica gel column chromatography.

[0052] 1H NMR (300MHz, DMSO) δ (ppm) 11.87 (s, 1H), 11.03 (s, 1H), 9.77 (s, 1H), 8.41 (d, J = 4.7Hz, 2H), 8.29-8.18 (m ,1H),7.86-7.71(m,3H),7.62(s,1H),7.47(d,J=8.4Hz,2H),7.41-7.31(m,1H),5.14(dd,J=13.2,5.1Hz ,1H),4.36(d,J=3.8Hz,2H),3.64(s,2H),3.22(dd,J=30.0,5.5Hz,6H),3.06-2.83(m,2H),2.56(d,J=6. 4Hz, 5H), 2.35 (t, J = 7.3Hz, 3H), 2.01 (d, J = 16.3Hz, 1H), 1.58 (dp, J = 22.4, 7.2Hz, 4H), 1.38-1.14 (m, 8H).

[0053] HRMS (ESI) (m / z): calculated value C 40 H 45 N 9 O 6 [M+H], 748.3526; found 748.3627.

[0054] Example 3: Preparation of N-(7-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)amino)-7-oxoheptyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-3)

[0055]

[0056] 295 mg (1.2 mmol) of Boc-7-aminoheptanoic acid and 495 mg (1.3 mmol) of HATU were dissolved in 5 ml of DMF, 523 μl (3 mmol) of DIPEA were added, and after stirring for 10 min, 260 mg (1 mmol) of lenalidomide was added, and the mixture was stirred at room temperature overnight. The reaction solution was extracted with EA and water, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-3 (370 mg, yield 76%) was obtained by column chromatography.

[0057] Compound A-3 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1, V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, and stirred until dissolved. 0.2 mmol HATU and 0.15 mmol compound C were added, and stirred at room temperature overnight. 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH. Compound I-3 (63 mg, yield 55.1%) was obtained by silica gel column chromatography.

[0058] 1 H NMR (400MHz, DMSO) δ (ppm) 11.88 (s, 1H), 11.03 (s, 1H), 9.81 (s, 1H), 8.41 (q, J = 4.9, 3.4Hz, 2H), 8.25 (d, J = 2.9Hz, 1H), 7.81 (t, J = 8.5Hz, 2 H),7.75(d,J=1.3Hz,1H),7.63(d,J=1.8Hz,1H),7.52-7.45(m,2H),7.38(dd,J=8.9,2.8Hz,1H),5.14(dd,J=13.3,5.1Hz,1H),4.43-4.30( m,2H),3.64(s,2H),3.32(d,J=5.9Hz,3H),3.25(q,J=6.7Hz,2H),3.17(d,J=5.1Hz,1H),2.97-2.84(m,1H),2.61(d,J=3.9Hz,1H),2.57-2 .52(m,5H),2.41-2.28(m,3H),2.01(ddd,J=12.5,6.4,3.8Hz,1H),1.56(dp,J=32.6,6.9Hz,4H),1.34-1.21(m,8H),1.17(t,J=7.4Hz,3H).

[0059] HRMS (ESI) (m / z): calculated value C 41 H 47 N 9 O 6 [M+H], 762.3683; found 762.3781.

[0060] Example 4: Preparation of N-(8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)amino)-8-oxooctyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-4)

[0061]

[0062] 312 mg (1.2 mmol) of Boc-8-aminocaprylic acid and 495 mg (1.3 mmol) of HATU were dissolved in 5 ml of DMF, 523 μl (3 mmol) of DIPEA were added, and after stirring for 10 min, 260 mg (1 mmol) of lenalidomide was added, and the mixture was stirred at room temperature overnight. The reaction solution was extracted with EA and water, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-4 (360 mg, yield 72%) was obtained by column chromatography.

[0063] Compound A-4 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, and stirred until dissolved. 0.2 mmol HATU and 0.15 mmol compound C were added, and stirred at room temperature overnight. 12 ml EA was added and stirred for 15 min, filtered, and the obtained solid was dissolved in DCM / MeOH. Compound I-4 (63 mg, yield 48.9%) was obtained by silica gel column chromatography.

[0064] 1 H NMR (300MHz, DMSO) δ (ppm) 11.88 (s, 1H), 11.03 (s, 1H), 9.77 (s, 1H), 8.41 (d, J = 1.7Hz, 2H), 8.26 (d, J = 2.8Hz, 1 H),7.86-7.77(m,2H),7.75(s,1H),7.63(s,1H),7.53-7.46(m,2H),7.39(dd,J=9.3,2.5Hz,1H),5.14(dd,J=13 .3,5.0Hz,1H),4.44-4.26(m,2H),3.65(s,2H),3.25(d,J=6.7Hz,2H),2.99-2.84(m,1H),2.65-2.52(m,5H),2. 34(t,J=7.5Hz,3H),2.01(d,J=13.0Hz,1H),1.54(d,J=29.0Hz,4H),1.38-1.21(m,12H),1.18(t,J=7.4Hz,4H).

[0065] HRMS (ESI) (m / z): calculated value C 42 H 49 N 9 O 6 [M+H], 776.3839; found 776.3961.

[0066] Example 5: Preparation of N-(9-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)amino)-9-oxononyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-5)

[0067]

[0068] 328 mg (1.2 mmol) of Boc-9-aminononanoic acid and 495 mg (1.3 mmol) of HATU were dissolved in 5 ml of DMF, 523 μl (3 mmol) of DIPEA were added, and after stirring for 10 min, 260 mg (1 mmol) of lenalidomide was added, and the mixture was stirred at room temperature overnight. The reaction solution was extracted with EA and water, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-5 (405 mg, yield 79%) was obtained by column chromatography.

[0069] Compound A-5 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-5 (63 mg, yield 48.9%) was obtained by silica gel column chromatography.

[0070] 1H NMR (300MHz, DMSO) δ (ppm) 11.89 (s, 1H), 11.04 (s, 1H), 9.81 (s, 1H), 8.41 (d, J = 5.1Hz, 2H), 8.26 (d, J = 2.8Hz, 1H), 7.85- 7.78(m,2H),7.75(s,1H),7.63(s,1H),7.52-7.43(m,2H),7.38(dd,J=8.9,2.9Hz,1H),5.14(dd,J=13.2,5.1Hz,1H),4.4 4-4.27(m,2H),3.68-3.60(m,2H),3.34-3.07(m,6H),2.92(ddd,J=17.9,13.4,5.3Hz,1H),2.66-2.52(m,5H),2.34(dd,J =10.3, 5.0Hz, 3H), 2.08-1.97 (m, 1H), 1.54 (dt, J = 28.7, 6.9Hz, 4H), 1.27 (td, J = 6.4, 4.1Hz, 12H), 1.17 (t, J = 7.4Hz, 3H).

[0071] HRMS (ESI) (m / z): calculated value C 43 H 51 N 9 O 6 [M+H], 790.3996; found 790.4077.

[0072] Example 6: Preparation of N-(10-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)amino)-10-oxodecanyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-6)

[0073]

[0074] 344 mg (1.2 mmol) of Boc-10-aminodecanoic acid and 495 mg (1.3 mmol) of HATU were dissolved in 5 ml of DMF, 523 μl (3 mmol) of DIPEA were added, and after stirring for 10 min, 260 mg (1 mmol) of lenalidomide was added, and the mixture was stirred at room temperature overnight. The reaction solution was extracted with EA and water, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-6 (403 mg, yield 76.3%) was obtained by column chromatography.

[0075] Compound A-5 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-6 (55 mg, yield 45.6%) was obtained by silica gel column chromatography.

[0076] 1 H NMR (300MHz, DMSO) δ (ppm) 11.88 (s, 1H), 11.04 (s, 1H), 9.78 (s, 1H), 8.39 (t, J = 4.5Hz, 2H), 8.26 (d, J = 2.8Hz, 1H), 7.87-7.78 (m, 2H ),7.75(s,1H),7.62(d,J=1.9Hz,1H),7.53-7.43(m,2H),7.38(dd,J=8.9,2.8Hz,1H),5.15(dd,J=13.2,5.1Hz,1H),4.44-4.28(m,2 H),3.64(s,2H),3.34-3.28(m,3H),3.24(q,J=6.6Hz,2H),3.17(d,J=5.2Hz,1H),2.92(ddd,J=17.9,13.4,5.2Hz,1H),2.65-2.51(m ,6H),2.34(dd,J=9.5,5.7Hz,3H),2.07-1.97(m,1H),1.53(dt,J=28.9,7.0Hz,4H),1.24(d,J=14.0Hz,12H),1.17(t,J=7.4Hz,3H).

[0077] HRMS (ESI) (m / z): calculated value C 44 H 53 N 9 O 6 [M+H],804.4152; measured value 804.4319.

[0078] Example 7: Preparation of N-(11-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)amino)-11-oxoundecyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-7)

[0079]

[0080] 362 mg (1.2 mmol) of Boc-11-aminoundecanoic acid and 495 mg (1.3 mmol) of HATU were dissolved in 5 ml of DMF, 523 μl (3 mmol) of DIPEA were added, and after stirring for 10 min, 260 mg (1 mmol) of lenalidomide was added, and the mixture was stirred at room temperature overnight. The reaction solution was extracted with EA and water, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-7 (460 mg, yield 84.8%) was obtained by column chromatography.

[0081] Compound A-5 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-7 (30 mg, yield 24.5%) was obtained by silica gel column chromatography.

[0082] 1 H NMR(300MHz,DMSO)δ(ppm)11.88(s,1H),11.04(s,1H),9.77(s,1H),8.40(s,2H),8.26(s,1H),7.82(d ,J=9.3Hz,2H),7.75(s,1H),7.63(s,1H),7.48(d,J=7.4Hz,2H),7.38(d,J=9.2Hz,1H),5.15(d,J=13. 1Hz,1H),4.44-4.27(m,2H),3.65(s,2H),3.30-3.08(m,3H),2.92(t,J=15.0Hz,1H),2.63(s,1H),2.5 6(d,J=6.7Hz,4H),2.34(t,J=7.6Hz,3H),1.99(d,J=2.8Hz,1H),1.53(d,J=32.2Hz,4H),1.25(s,20H).

[0083] HRMS (ESI) (m / z): calculated value C 45 H 55 N 9 O 6 [M+H], 818.4309; found 818.4346.

[0084] Example 8: Preparation of N-(12-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-4-yl)amino)-12-oxododecyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-8)

[0085]

[0086] 379 mg (1.2 mmol) of Boc-12-aminododecanoic acid and 495 mg (1.3 mmol) of HATU were dissolved in 5 ml of DMF, 523 μl (3 mmol) of DIPEA were added, and after stirring for 10 min, 260 mg (1 mmol) of lenalidomide was added, and the mixture was stirred at room temperature overnight. The reaction solution was extracted with EA and water, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-8 (436 mg, yield 78.3%) was obtained by column chromatography.

[0087] Compound A-5 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-8 (36 mg, yield 28.8%) was obtained by silica gel column chromatography.

[0088] 1H NMR (300MHz, DMSO) δ (ppm) 11.87 (s, 1H), 11.04 (s, 1H), 9.79 (s, 1H), 8.44-8.36 (m, 2H), 8.25 (d, J = 2.8Hz, 1H), 7 .86-7.78(m,2H),7.74(s,1H),7.62(d,J=1.8Hz,1H),7.52-7.46(m,2H),7.37(dd,J=8.9,2.8Hz,1H),5.14(dd,J =13.3,5.1Hz,1H),4.43-4.28(m,2H),3.64(s,2H),3.27(dt,J=31.3,5.7Hz,6H),2.99-2.84(m,2H),2.65-2.52( m,6H),2.38-2.30(m,3H),2.02(ddd,J=11.0,5.9,3.4Hz,1H),1.53(dt,J=31.6,6.9Hz,4H),1.27-1.14(m,18H).

[0089] HRMS (ESI) (m / z): calculated value C 46 H 57 N 9 O 6 [M+H], 832.4465; found 832.4561.

[0090] Example 9: Preparation of N-(4-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)amino)-4-oxobutyl)-5-(4-(7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-9)

[0091]

[0092] Dissolve 273 mg (1 mmol) of 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione in 5.6 ml of DMSO, heat to 160°C, add 523 μl (3 mmol) of DIPEA and 168 mg (1.05 mmol) of N-tert-butoxycarbonyl-1,2-ethylenediamine, stir for 10 min, extract the reaction solution with EA and water, combine the organic layers, wash with saturated brine, dry over anhydrous sodium sulfate, filter, concentrate the organic phase, and obtain compound A-9 (375 mg, yield 91%) by column chromatography.

[0093] Compound A-9 (0.5 mmol) was dissolved in 5 ml DCM / TFA (2:1, V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 3 ml DMF and 350 μl (2 mmol) DIPEA were added, stirred until dissolved, 0.6 mmol HATU and 0.6 mmol N-Boc-aminobutyric acid were added, stirred at room temperature overnight, the reaction solution was extracted with EA and water, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-10 (176 mg, yield 70.1%) was obtained by column chromatography.

[0094] Compound A-10 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the obtained solid was dissolved in DCM / MeOH, and compound I-9 (45 mg, yield 38.6%) was obtained by silica gel column chromatography.

[0095] 1 H NMR (300MHz, DMSO) δ (ppm) 12.18 (s, 1H), 11.10 (s, 1H), 8.73-8.44 (m, 2H), 8.30 (s, 1H), 8.11 (t, J = 5.5Hz, 1H), 7.90-7.75 (m, 3 H),7.62-7.52(m,1H),7.46(d,J=8.6Hz,1H),7.17(d,J=8.6Hz,1H),7.01(d,J=7.0Hz,1H),6.74(t,J=6.1Hz,1H),5.04(dd,J=1 2.6,5.3Hz,1H),4.51(s,1H),4.02(dd,J=14.1,7.1Hz,1H),3.64-3.52(m,1H),3.29-3.02(m,7H),2.94-2.79(m,2H),2.73(s,1 H), 2.56 (dd, J = 14.9, 7.2Hz, 4H), 2.13-1.95 (m, 3H), 1.73 (p, J = 7.1Hz, 2H), 1.27 (dd, J = 9.4, 6.5Hz, 2H), 1.19 (t, J = 7.4Hz, 3H).

[0096] HRMS (ESI) (m / z): calculated value C 40 H 44 N 10 O 7[M+H], 777.3428; found 777.3600.

[0097] Example 10: Preparation of N-(5-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)amino)-5-oxopentyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-10)

[0098]

[0099] Compound A-9 (0.5 mmol) was dissolved in 5 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was dried, 3 ml DMF and 350 μl (2 mmol) DIPEA were added, stirred until dissolved, 0.6 mmol HATU and 0.6 mmol Boc-5-aminopentanoic acid were added, stirred at room temperature overnight, the reaction solution was extracted with EA and water, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-11 (154 mg, yield 59.7%) was obtained by column chromatography.

[0100] Compound A-11 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-10 (52 mg, yield 43.8%) was obtained by silica gel column chromatography.

[0101] 1H NMR (300MHz, DMSO) δ (ppm) 11.89 (s, 1H), 11.10 (s, 1H), 8.42 (d, J = 5.2Hz, 2H), 8.26 (d, J = 2.7Hz, 1H), 8.05 (t, J = 5.6Hz, 1H), 7. 82(d,J=8.7Hz,1H),7.75(s,1H),7.64(s,1H),7.56(t,J=7.8Hz,1H),7.43-7.34(m,1H),7.16(d,J=8.7Hz,1H),7.00(d,J=7.1 Hz,1H),6.77-6.67(m,1H),5.05(dd,J=12.8,5.3Hz,1H),3.66(s,2H),3.23(d,J=6.2Hz,4H),2.90-2.80(m,1H),2.61-2.52(m ,6H),2.04(dt,J=18.9,6.1Hz,3H),1.91(s,1H),1.48(s,4H),1.37-1.27(m,2H),1.23(d,J=8.8Hz,3H),1.18(t,J=7.4Hz,4H).

[0102] HRMS (ESI) (m / z): calculated value C 41 H 46 N 10 O 7 [M+H], 791.3584; found 791.3558.

[0103] Example 11: Preparation of N-(6-((2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)amino)-6-oxohexyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-11)

[0104]

[0105] Compound A-9 (0.5 mmol) was dissolved in 5 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 3 ml DMF and 350 μl (2 mmol) DIPEA were added, stirred until dissolved, 0.6 mmol HATU and 0.6 mmol Boc-6-aminohexanoic acid were added, stirred at room temperature overnight, the reaction solution was extracted with EA and water, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-12 (179 mg, yield 67.7%) was obtained by column chromatography.

[0106] Compound A-12 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-11 (28 mg, yield 23.2%) was obtained by silica gel column chromatography.

[0107] 1 H NMR (400MHz, DMSO) δ (ppm) 11.87 (s, 1H), 11.10 (s, 1H), 8.38 (dd, J = 12.3, 6.3Hz, 2H), 8.23 ​​(d, J = 2.9Hz, 1H), 8.02 (t, J = 5.6Hz, 1H), 7.81 (d, J = 8.8Hz, 1H) ,7.75(d,J=1.4Hz,1H),7.64-7.61(m,1H),7.57(dd,J=8.6,7.0Hz,1H),7.37 (dd,J=8.9,2.9Hz,1H),7.15(d,J=8.6Hz,1H),7.01(d,J=7.0Hz,1H),6.71(t ,J=6.2Hz,1H),5.06(dd,J=12.9,5.4Hz,1H),3.64(s,2H),3.32(q,J=5.9,5. 2Hz, 4H), 3.23 (dq, J=11.3, 6.1, 4.7Hz, 4H), 3.16 (d, J=5.1Hz, 1H), 2.88 (ddd ,J=17.3,14.0,5.4Hz,1H),2.61-2.52(m,6H),2.09-1.97(m,3H),1.49(h,J= 7.2Hz, 4H), 1.29 (d, J = 2.7Hz, 1H), 1.28-1.20 (m, 6H), 1.18 (t, J = 7.4Hz, 3H).

[0108] HRMS (ESI) (m / z): calculated value C 42 H 48 N 10 O 7 [M+H], 805.3741; found 805.3845.

[0109] Example 12: Preparation of N-(7-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)amino)-7-oxoheptyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-12)

[0110]

[0111] Compound A-9 (0.5 mmol) was dissolved in 5 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 3 ml DMF and 350 μl (2 mmol) DIPEA were added, stirred until dissolved, 0.6 mmol HATU and 0.6 mmol Boc-7-aminoheptanoic acid were added, stirred at room temperature overnight, the reaction solution was extracted with EA and water, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-13 (163 mg, yield 60.0%) was obtained by column chromatography.

[0112] Compound A-13 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-12 (36 mg, yield 29.3%) was obtained by silica gel column chromatography.

[0113] 1H NMR (300MHz, DMSO) δ (ppm) 11.90 (s, 1H), 11.11 (s, 1H), 8.44-8.35 (m, 2H), 8.26 (d, J = 2.8Hz, 1H), 8.03 (t, J = 5.5Hz, 1H), 7.82 (d, J = 8.8Hz, 1H), 7 .75(s,1H),7.64(s,1H),7.57(dd,J=8.6,7.1Hz,1H),7.39(dd,J=8.9,2.9Hz,1H),7.16(d,J=8.6Hz,1H),7.01(d,J=7.0Hz,1H),6.71(t,J=6.0H z,1H),5.05(dd,J=12.8,5.4Hz,1H),3.75-3.56(m,2H),3.27-3.07(m,5H),2.88(ddd,J=16.9,13.8,5.4Hz,1H),2.66-2.52(m,6H),2.03(t,J=7 .3Hz,3H),1.46(t,J=7.0Hz,4H),1.29(d,J=2.4Hz,2H),1.28-1.26(m,3H),1.24(d,J=3.0Hz,3H),1.22(d,J=2.0Hz,5H),1.18(t,J=7.4Hz,3H).

[0114] HRMS (ESI) (m / z): calculated value C 43 H 50 N 10 O 7 [M+H], 819.3897; found 819.4064.

[0115] Example 13: Preparation of N-(8-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)amino)-8-oxooctyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-13)

[0116]

[0117] Compound A-9 (0.5 mmol) was dissolved in 5 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was dried, 3 ml DMF and 350 μl (2 mmol) DIPEA were added, stirred until dissolved, 0.6 mmol HATU and 0.6 mmol Boc-8-aminocaprylic acid were added, stirred at room temperature overnight, the reaction solution was extracted with EA and water, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-14 (201 mg, yield 72.0%) was obtained by column chromatography.

[0118] Compound A-14 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-13 (52 mg, yield 41.6%) was obtained by silica gel column chromatography.

[0119] 1 H NMR (300MHz, DMSO) δ (ppm) 11.87 (s, 1H), 11.11 (s, 1H), 8.44-8.32 (m, 2H), 8.26 (d, J = 2.9Hz, 1H), 8.01 (t, J = 5.5Hz, 1H), 7.82 (d ,J=8.8Hz,1H),7.75(d,J=1.5Hz,1H),7.63(d,J=1.8Hz,1H),7.56(dd,J=8.6,7.1Hz,1H),7.38(dd,J=8.9,2.7Hz,1H),7.16(d,J =8.6Hz,1H),7.02(d,J=7.0Hz,1H),6.71(t,J=6.1Hz,1H),5.05(dd,J=12.8,5.4Hz,1H),3.70-3.60(m,2H),3.20(dt,J=17.2,5. 6Hz,5H),2.95-2.81(m,1H),2.63-2.52(m,7H),2.02(q,J=6.8,6.3Hz,3H),1.54-1.40(m,4H),1.34(s,1H),1.30-1.13(m,14H).

[0120] HRMS (ESI) (m / z): calculated value C 44 H 52 N 10 O 7[M+H], 833.4054; found 833.4128.

[0121] Example 14: Preparation of N-(8-((3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)propyl)amino)-8-oxooctyl)-5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-14)

[0122]

[0123] Dissolve 273 mg (1 mmol) of 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione in 5.6 ml of DMSO, heat to 160°C, add 523 μl (3 mmol) of DIPEA and 183 mg (1.05 mmol) of N-tert-butyloxycarbonyl-1,3-propylenediamine, stir for 10 min, extract the reaction solution with EA and water, combine the organic layers, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the organic phase, and obtain compound A-15 (400 mg, yield 93%) by column chromatography.

[0124] Compound A-15 (0.5 mmol) was dissolved in 5 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 3 ml DMF and 350 μl (2 mmol) DIPEA were added, stirred until dissolved, 0.6 mmol HATU and 0.6 mmol Boc-8-aminocaprylic acid were added, stirred at room temperature overnight, the reaction solution was extracted with EA and water, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-16 (216 mg, yield 75.6%) was obtained by column chromatography.

[0125] Compound A-16 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, stirred until dissolved, 0.2 mmol HATU and 0.15 mmol compound C were added, stirred at room temperature overnight, 12 ml EA was added and stirred for 15 min, filtered, and the resulting solid was dissolved in DCM / MeOH, and compound I-14 (55 mg, yield 43.3%) was obtained by silica gel column chromatography.

[0126] 1H NMR (300MHz, DMSO) δ (ppm) 11.88 (s, 1H), 11.11 (s, 1H), 8.39 (dd, J = 7.1, 4.1Hz, 2H), 8.25 (d, J = 2.8Hz, 1H), 7.90-7.79 (m, 2H), 7 .74(s,1H),7.63(s,1H),7.56(dd,J=8.6,7.1Hz,1H),7.38(dd,J=8.9,2.8Hz,1H),7.03(dd,J=15.7,7.8Hz,2H),6.67(t,J=6.1H z,1H),5.05(dd,J=12.7,5.4Hz,1H),3.70-3.58(m,2H),3.34-3.19(m,7H),3.12(dq,J=12.4,6.3,5.6Hz,2H),2.88(ddd,J=17. 1,13.8,5.3Hz,1H),2.63-2.52(m,6H),2.05(t,J=7.4Hz,3H),1.65(t,J=6.6Hz,2H),1.48(h,J=6.8Hz,4H),1.31-1.12(m,12H).

[0127] HRMS (ESI) (m / z): calculated value C 45 H 54 N 10 O 7 [M+H], 847.4210; found 847.4310.

[0128] Example 15: Preparation of N-(8-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)butyl)amino)-8-oxooctyl)-5-(4-(7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-15)

[0129]

[0130] Dissolve 273 mg (1 mmol) of 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione in 5.6 ml of DMSO, heat to 160°C, add 523 μl (3 mmol) of DIPEA and 198 mg (1.05 mmol) of N-tert-butyloxycarbonyl-1,4-butanediamine, stir for 10 min, extract the reaction solution with EA and water, combine the organic layers, wash with saturated brine, dry over anhydrous sodium sulfate, filter, concentrate the organic phase, and obtain compound A-17 (397 mg, yield 89.3%) by column chromatography.

[0131] Compound A-17 (0.5 mmol) was dissolved in 5 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 3 ml DMF and 350 μl (2 mmol) DIPEA were added, stirred until dissolved, 0.6 mmol HATU and 0.6 mmol Boc-8-aminocaprylic acid were added, stirred at room temperature overnight, the reaction solution was extracted with EA and water, the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-18 (176 mg, yield 60.1%) was obtained by column chromatography.

[0132] Compound A-18 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, and stirred until dissolved. 0.2 mmol HATU and 0.15 mmol compound C were added, and stirred at room temperature overnight. 12 ml EA was added and stirred for 15 min, filtered, and the obtained solid was dissolved in DCM / MeOH. Compound I-15 (55 mg, yield 43.3%) was obtained by silica gel column chromatography.

[0133] 1 H NMR (300MHz, DMSO) δ (ppm) 11.88 (s, 1H), 11.11 (s, 1H), 8.39 (dd, J = 7.8, 4.1Hz, 2H), 8.25 (d, J = 2.8Hz, 1H), 7.84-7.72 (m, 3H), 7.6 3(d,J=1.9Hz,1H),7.55(dd,J=8.6,7.0Hz,1H),7.37(dd,J=8.9,2.8Hz,1H),7.08(d,J=8.6Hz,1H),7.01(d,J=7.0Hz,1H),6.54(t ,J=6.0Hz,1H),5.05(dd,J=12.8,5.4Hz,1H),3.64(s,2H),3.34-3.18(m,7H),3.06(q,J=6.4Hz,2H),2.88(ddd,J=17.0,13.8,5.4 Hz,1H),2.63-2.52(m,6H),2.02(dd,J=9.8,4.8Hz,3H),1.49(dq,J=19.6,6.9,6.0Hz,8H),1.21(ddd,J=17.4,12.4,7.1Hz,13H).

[0134] HRMS (ESI) (m / z): calculated value C 46 H 56 N 10 O 7[M+H], 861.4367; found 861.4460.

[0135] Example 16: Preparation of N-(8-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)butyl)amino)-8-oxooctyl)-5-(4-(7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinamide (Compound I-16)

[0136]

[0137] Dissolve 273 mg (1 mmol) of 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione in 5.6 ml of DMSO, heat to 160°C, add 523 μl (3 mmol) of DIPEA and 210 mg (1.05 mmol) of 3-tert-butoxycarbonylaminopiperidine, stir for 10 min, extract the reaction solution with EA and water, combine the organic layers, wash with saturated brine, dry over anhydrous sodium sulfate, filter, concentrate the organic phase, and obtain compound A-21 (375 mg, yield 82%) by column chromatography.

[0138] Compound A-21 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, and stirred until dissolved. 0.2 mmol HATU and 0.15 mmol compound C were added, and stirred at room temperature overnight. 12 ml EA was added and stirred for 15 min, filtered, and the obtained solid was dissolved in DCM / MeOH. Compound I-16 (62 mg, yield 57.4%) was obtained by silica gel column chromatography.

[0139] 1H NMR (300MHz, DMSO) δ (ppm) 11.87 (s, 1H), 11.09 (d, J = 1.9Hz, 1H), 8.45-8.37 (m, 2H), 8.25 (t, J = 2.3Hz, 1H), 7.85 (d, J = 8.8Hz, 1H ),7.74(d,J=1.5Hz,1H),7.69(dd,J=8.5,7.1Hz,1H),7.62(d,J=1.8Hz,1H),7.39(dd,J=8.8,3.2Hz,2H),7.34(d,J=7.1Hz,1H) ,5.14-5.04(m,1H),4.08(s,1H),3.65(s,2H),3.57(d,J=7.6Hz,2H),3.08(dq,J=20.1,10.9,10.4Hz,2H),2.95-2.79(m,1H),2 .62-2.52(m,7H),2.09-1.94(m,1H),1.79(dd,J=32.5,14.4Hz,4H),1.34(d,J=4.4Hz,1H),1.21(s,4H),1.17(t,J=7.4Hz,3H).

[0140] HRMS (ESI) (m / z): calculated value C 39 H 41 N 9 O 6 [M+H], 732.3213; found 732.3394.

[0141] Example 17: Preparation of 2-(2,6-dioxopiperidin-3-yl)-4-(9-(5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthopyridin-3-yl)methyl)piperazin-1-yl)picolinyl)-3,9-diazaspiro[5.5]undecane-3-yl)isoindoline-1,3-dione (Compound I-17)

[0142]

[0143] 137 mg (0.5 mmol) of 2-(2,6-dioxo-piperidin-3-yl)-4-fluoro-isoindole-1,3-dione was dissolved in 2.8 ml of DMSO, and the temperature was raised to 160°C. 262 μl (1.5 mmol) of DIPEA and 122 mg (1.05 mmol) of tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate were added, and the mixture was stirred for 10 min. The reaction solution was extracted with EA and water, and the organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. Compound A-20 (215 mg, yield 84.1%) was obtained by column chromatography.

[0144] Compound A-20 (0.18 mmol) was dissolved in 1.8 ml DCM / TFA (2:1 V / V), reacted at room temperature for 1 h, the solvent was spin-dried, 1.5 ml DMF and 126 μl (0.72 mmol) DIPEA were added, and stirred until dissolved. 0.2 mmol HATU and 0.15 mmol compound C were added, and stirred at room temperature overnight. 12 ml EA was added and stirred for 15 min, filtered, and the obtained solid was dissolved in DCM / MeOH. Compound I-17 (62 mg, yield 52.6%) was obtained by silica gel column chromatography.

[0145] 1 H NMR (300MHz, DMSO) δ (ppm) 11.87 (s, 1H), 11.10 (s, 1H), 8.40 (d, J = 1.8Hz, 1H), 8.25 (d, J = 2.8Hz, 1H), 7.74 (s, 1H), 7.66(dd,J=8.4,7.1Hz,1H),7.62(d,J=1.8Hz,1H),7.46(d,J=8.7Hz,1H),7.40-7.35(m,1H),7.32(dd,J=7.8,4.7 Hz,2H),5.08(dd,J=12.6,5.4Hz,1H),3.59(d,J=26.2Hz,6H),3.28(s,7H),2.93-2.78(m,1H),2.63-2.51(m,7H), 2.03(d,J=5.4Hz,1H),1.66(s,4H),1.50(d,J=16.1Hz,4H),1.34(s,1H),1.23-1.18(m,4H),1.15(d,J=1.5Hz,1H).

[0146] HRMS (ESI) (m / z): calculated value C 43 H 47 N 9 O 6 [M+H], 786.3683; found 786.3841.

[0147] Example 18 PARP1 protein degradation experiment in MDA-MB-231 cells

[0148] MDA-MB-231 cells (ATCC) were cultured in DMEM complete medium at 37°C and 5% CO. 2 DMEM medium was purchased from Gibco and contained 10% FBS (ExCell Bio) and 1% Penicillin-Streptomycin (Gibco). Cells in the logarithmic growth phase were collected and the cell concentration was adjusted to 1.5×105 / ml, add 1ml to each well of a 12-well cell culture plate. After 24h, observe the cell growth status and whether it is evenly distributed under a microscope. The test compound is prepared into a 10mM stock solution using DMSO, and then diluted with DMEM basal culture medium to solutions of different concentrations (0.3nM-1000nM). Discard the original culture medium in the well plate, add different concentrations of drugs respectively, and the volume of solution in each well is 1ml. After 24h of administration, the plate is collected, the original culture medium is discarded, and after washing once with pre-cooled PBS, 1ml PBS is added, and the cells are gently blown off with a gun. This step is completed on ice. The obtained cell suspension was transferred to a 1.5 ml centrifuge tube and centrifuged at 3500 RPM at 4°C for five minutes to obtain a cell pellet. The upper PBS layer was discarded and 60 ul of RIPA lysis buffer containing protease inhibitors (100×) and phosphatase inhibitors (50×) was added (protease inhibitors and phosphatase inhibitors were purchased from Beyotime). After sonication, the tube was lysed on ice for 20 minutes, and then centrifuged at 4°C, 13000 g for 10 minutes. The supernatant was collected to obtain the protein.

[0149] The protein concentration was determined by BCA method, and the protein concentration was normalized and then added to loading buffer (Bio-Rad), vortexed and boiled at 100°C for 8 min to obtain protein samples for subsequent Western blot detection. Western blot experiments used 8% separation gel, PARP1 antibody was purchased from CST (PARP (46D11) Rabbit mAb #9532), and internal reference Anti-alpha Tubulin was purchased from Hangzhou Huaan Biotechnology Co., Ltd. (JM73-24).

[0150] The experimental results are shown in Table 1:

[0151] Table 1 PARP1 degradation level in MDA-MB-231 cells

[0152] serial number DC50(nM) <![CDATA[D max ]]> I-1 - 5％-50％ I-2 - 5％-50％ I-3 - 5％-50％ I-4 >10 5％-80％ I-5 - 5％-50％ I-6 2.951 5％-80％ I-7 7.1 >80％ I-8 3.299 >80％ I-9 >10 5％-80％ I-10 - 5％-50％ I-11 >10 5％-80％ I-12 2.869 >85％ I-13 <1 >85％ I-14 >10 5％-80％ I-15 >10 5％-80％ I-16 - 5％-50％ I-17 - 5％-50％

[0153] DC50 represents the drug concentration required to degrade 50% of the protein. max Represents the maximum protein degradation rate that can be caused.

[0154] As described above, although the present invention has been shown and described with reference to specific preferred embodiments, it should not be construed as limiting the present invention itself. Various changes in form and details may be made without departing from the spirit and scope of the present invention as defined in the appended claims.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof: in, L is a linker selected from a C5-C12 aliphatic chain, a saturated heterocycle, or an alkyl chain containing 2-4 N atoms; D is selected from:

2. The compound according to claim 1, characterized in that L is selected from:

3. The compound according to claim 1, characterized in that: The pharmaceutically acceptable salts include acid addition salts formed between the compound of formula I and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid; and also include acid salts formed between the compound of formula I and inorganic bases.

4. The compound according to claim 1, characterized in that Selected from:

5. A method for preparing the compound of general formula I according to claim 1, characterized in that: The following steps are involved: (1) Compound A undergoes a nucleophilic reaction with an aliphatic hydrocarbon chain, a heteroaromatic ring structure compound, or an alkyl chain compound containing 2 to 4 N atoms, or undergoes a condensation reaction in the presence of a condensation agent to obtain Compound B; (2) Compound B is subjected to acid hydrolysis to remove the Boc protecting group, and then undergoes a condensation reaction with compound C in the presence of an amide condensation agent to generate a compound of formula I. wherein X is selected from H, O, and L and D are as defined in Formula I.

6. A pharmaceutical composition, characterized in that: It comprises the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

7. The pharmaceutical composition according to claim 6, characterized in that: The dosage form of the pharmaceutical composition is tablet, capsule, powder, syrup, liquid, suspension, lyophilized powder or injection.

8. Use of the compound according to any one of claims 1 to 4 in the preparation of a PARP1 degrader.

9. Use of the compound according to any one of claims 1 to 4 in the preparation of a drug for treating cancer.

10. The use according to claim 9, characterized in that: The cancer is triple-negative breast cancer.

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