医学以bcl2蛋白家族为靶点的抗肿瘤药物设计

1、会计学1医学以医学以bcl2蛋白家族为靶点的抗肿瘤药蛋白家族为靶点的抗肿瘤药物设计物设计2,Cell,144(5): 646-674Hallmarks of Cancer3Extrinsic and intrinsic pathways of apoptosisExpert Opin. Ther. Patents (2012) ,22(1):37-554The Bcl-2 Family Proteins with anti- or pro-apoptotic function Characteristics: (Often) a C-terminal transmembrane domain.

2、 Bcl-2 homology (BH) domains. Bcl-2 proteins form homo- and heterodimers via their BH-domains BH1, BH2: Predicted to form ion channels BH3:The ”suicide domain” regulates cell death BH4:Thought to confer anti-apoptotic activity.(Chan & Yu. (2004). Clin. Exp. Pharmacol. Physiol. 31: 119).Chipuk, J

3、. E，Molecular Cell ，2010，37(3): 299-3105Seminars in Oncology, 2004,30(Supplement 16): 133-142.6一、直接模型： 抑制剂代替促凋亡蛋白BAK/BAX，与抗凋亡蛋白Bcl-2/Bcl-xl结合，从而抑制后者，促进细胞凋亡二、间接模型 抑制剂替代特定的激活因子BH3-only蛋白，与抗调亡蛋白如BCL-2/BCL-xL结合，游离出来的BH3-only蛋白，激活BAK/BAX。Nature Reviews Drug Discovery 2008，7(12): 989-1000Clinical Cancer R

4、esearch 2009，15(4): 1126-1132.Oncogene ，2009，27(S1): S149-S157.7Structure basis for designing ligands89抗凋亡蛋白(BCL-2)是通过其表面疏水凹槽与促凋亡类(BIM)的 BH3区域结合发生相互作用，调节细胞正常的生理凋亡。BCL-2 family antagonists for cancer therapy. Nature Reviews Drug Discovery 2008，7(12): 989-1000Overlay of BCL-XLABT-737 complex with BCL-

5、xlLBIM BH3 complexBcl-2 and BIM BH3 1011. Structure of Bcl-xL-Bak Peptide Complex: Recognition Between Regulators of Apoptosis. Science 1997，275(5302): 983-986.Bcl-xL-Bim BH3Mcl-1Bim BH3Bcl-xL/BadProtein Science ,2000, 9:25282534125个疏水中心 (P1, P2, P3, P4和P5)4个氢键供体(D1, D2, D3和 D4)2个氢键受体(A1 and A2)Bcl-

6、2活性部位分布图在受体D1、D2、D3、A1位点出现比较频繁，与受体形成静电作用能提高亲和力与特异性D4，A2位点出现频率较小，能量贡献不大 BMC，2007， 15(19): 6407-6417Peptidomimetic approach mimic the a-helix of the BH3 peptidesABT-737, ABT-263 and their analogspan-active inhibitor of Bcl-2Selective Bcl-2 inhibitorsselective BCL-xL inhibitorSelective Mcl-1 inhibitors

7、14BakPeptidomimetic approach mimic the a-helix of the BH3 peptides JACS,2005,127(29): 10191-10196JACS,2005,127(15): 5,463-546815J Biol Chem. 2011 ,18; 286(11): 93829392 Organic Letters,2007, 9(19): 3733-3736.Bim BH3 a-helixJ. Med. Chem. 2012, 55, 107351074116173.Screen for second ligand2D 1H-15N HSQ

8、C谱中蛋白的H1或N15化学位移变化。18Science. 1996 Nov 29;274(5292):1531-4.Link ligands连接基团成功的关键是连接桥的长度及其性质（如刚性）Science 274, 1531-1534 (1996)19黑：HSQC of 15N-Bcl-xL红：HSQC of 15N-Bcl-xL withbiaryl acid（ 1 ）1的羧基能与R139结合，相当与Bcl-XlBak复合物中的D8320Site1:Tyr 101, Leu 108, Val 126 and Phe 14621黑: HSQC of 15N-Bcl-xL.红: HSQC of

9、 15N-Bcl-xL withbiaryl acid (1).绿: HSQC of 15N-Bcl-xL withnapthol (11).221123J Med Chem 49, 656-663 (2006)联苯替代Bak-BH3的V74、Leu78硝基苯替代Ile85酰基磺胺上O与R139接近，替代Bak的Asp83并且两个芳环与Y194，F97重叠，形成疏水性的 stacking作用Site 224A-38535831不幸的是，31在HSA存在情况下与Bcl-Xl结合大大下降。它能与HSA的domain III紧密结合。J. Med. Chem.2006,49,1165-118168倍

10、的失活Bcl-xL Ki 1nMBcl-xL Ki 10% HS:35nM25增加site1的极性ABT-737ABT-263 MW:974Pka10J. Med. Chem.2008,51,69026915Nature Reviews Drug Discovery 2008，7(12): 989-100026MW：813.43喹唑啉类JACS. 2006,128, 16206-16212J. Med. Chem.2011, 54,19141926J. Med. Chem.2012, 55, 46644682J. Med. Chem.2012, 55, 61496161J. Med. Chem

11、.2012, 55, 85028514J. Med. Chem.2013, 56, 304830674,5-二苯基吡唑类Bcl-2/Bcl-xL:Ki1 nMAOptimizationBM-95727282930Obatoclax Phase II AT-101Phase II/III早期研发的抗凋亡蛋白小分子抑制剂基本上都是pan-Bcl-2 antagonist包括三联苯类， HA-14-1及其类似物，棉酚及其衍生物，BH3I及其衍生物等 BH3I-1S1DCBL55Journal of Medicinal Chemistry,2010,53(9): 3465-3479Nature Rev

12、iews Drug Discovery ,2008,7(12): 989-100031 ABT-199 Selectively Kills BCL-2-Dependent Tumor Cells While Sparing Platelets (A) Navitoclax (ABT-263) binds with high affinity to both BCL-2 and BCL-XL. Because many tumors, particularly lymphoid malignancies, are addicted to BCL-2 for survival,.Cancer Ce

13、ll , 2013,23(2),139 - 141ABT-199：Selective Bcl-2 inhibitor32Nature medicine,2013,19(2):202-8BCL-2 Ki 0.010 nMBCL-XL Ki = 48 nMBCL-W Ki = 245 nM Phase I/II Phase I33化合物1与Bcl-2化合物2与Bcl-2化合物1与Bcl-2的晶体复合物中发现1与一个 Bcl-2结合时，另外一个Bcl-2的Trp30的吲哚环会插入P4位点与Asp103形成氢键。同时产生-stacking作用而Asp103相当于Bcl-xl的GLu96。34YC137

14、35Nature Chemical Biology 9, 390397 (2013)benzothiazole-hydrazone scaffoldP2P4苯并噻唑腙深入P2位点里面，并与Ser106,Leu108形成氢键作用羧基与Arg139结合，形成氢键作用苄胺在P4位点，并且紧密贴合。氨基与GLu96有静电作用WEHI-539替代Phe105,使得P2疏水口袋更深（ABT-737）。P2具有可塑性。P4BCL-XL：IC50= 1.1 nM选择性：500-fold over BCL-2 400-fold over BCL-W, MCL-1Arg139Phe105Tyr19536Glu96

15、 2.2 M0.77 M12化合物活性虽未提高，但晶体复合物发现，苯并噻唑腙深入P2位点里面，贴合紧密，并且与Ser106,Leu108形成氢键。酰基磺胺与Arg139的胍基相互作用 37优化38WEHI-539Marinopyrrole AJ Biol Chem. 2012 , 287：10224-10235J. Med. Chem.2013, 56, 1530WO2013052943A2Ki：0.32M10MUMI-77Ki：0.49M39404142The Bcl-2 Family Proteins with anti- or pro-apoptotic function Charact

16、eristics: (Often) a C-terminal transmembrane domain. Bcl-2 homology (BH) domains. Bcl-2 proteins form homo- and heterodimers via their BH-domains BH1, BH2: Predicted to form ion channels BH3:The ”suicide domain” regulates cell death BH4:Thought to confer anti-apoptotic activity.(Chan & Yu. (2004). Clin. Exp. Pharmacol. Physiol. 31: 119).Chipuk, J. E，Molecular Cell ，2010，37(3): 299-31043Structure basis for designing ligands44BakPeptidomimetic approach mimic the a-helix of the BH3 peptides JACS,2005,127