WO2021116929A1 - Composition comprising the bh3-mimetic abt-199 (venetoclax) and at least one inhibitor of the nadph production - Google Patents
Composition comprising the bh3-mimetic abt-199 (venetoclax) and at least one inhibitor of the nadph production Download PDFInfo
- Publication number
- WO2021116929A1 WO2021116929A1 PCT/IB2020/061684 IB2020061684W WO2021116929A1 WO 2021116929 A1 WO2021116929 A1 WO 2021116929A1 IB 2020061684 W IB2020061684 W IB 2020061684W WO 2021116929 A1 WO2021116929 A1 WO 2021116929A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- treatment
- abt
- lymphoma
- cell
- cell lymphoma
- Prior art date
Links
- LQBVNQSMGBZMKD-UHFFFAOYSA-N venetoclax Chemical compound C=1C=C(Cl)C=CC=1C=1CC(C)(C)CCC=1CN(CC1)CCN1C(C=C1OC=2C=C3C=CNC3=NC=2)=CC=C1C(=O)NS(=O)(=O)C(C=C1[N+]([O-])=O)=CC=C1NCC1CCOCC1 LQBVNQSMGBZMKD-UHFFFAOYSA-N 0.000 title claims abstract description 105
- 229960001183 venetoclax Drugs 0.000 title claims abstract description 104
- 239000003112 inhibitor Substances 0.000 title claims abstract description 24
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 17
- 239000000203 mixture Substances 0.000 title description 9
- ACFIXJIJDZMPPO-NCHANQSKSA-N nadph Chemical compound C1=CCC(C(=O)N)=CN1[C@@H]1[C@@H](O)[C@@H](O)[C@H](COP(O)(=O)OP(O)(=O)OC[C@H]2[C@@H]([C@H](OP(O)(O)=O)[C@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 ACFIXJIJDZMPPO-NCHANQSKSA-N 0.000 title 1
- 208000031671 Large B-Cell Diffuse Lymphoma Diseases 0.000 claims abstract description 80
- 206010012818 diffuse large B-cell lymphoma Diseases 0.000 claims abstract description 80
- 238000011282 treatment Methods 0.000 claims abstract description 63
- 208000031261 Acute myeloid leukaemia Diseases 0.000 claims abstract description 55
- FMGSKLZLMKYGDP-USOAJAOKSA-N dehydroepiandrosterone Chemical compound C1[C@@H](O)CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CC=C21 FMGSKLZLMKYGDP-USOAJAOKSA-N 0.000 claims abstract description 48
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims abstract description 22
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 22
- 150000001875 compounds Chemical class 0.000 claims abstract description 21
- 206010000830 Acute leukaemia Diseases 0.000 claims abstract description 18
- 208000028564 B-cell non-Hodgkin lymphoma Diseases 0.000 claims abstract description 18
- 230000002438 mitochondrial effect Effects 0.000 claims abstract description 17
- 208000025205 Mantle-Cell Lymphoma Diseases 0.000 claims abstract description 15
- 230000037361 pathway Effects 0.000 claims abstract description 15
- 208000036170 B-Cell Marginal Zone Lymphoma Diseases 0.000 claims abstract description 14
- 229940014144 folate Drugs 0.000 claims abstract description 14
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims abstract description 14
- 235000019152 folic acid Nutrition 0.000 claims abstract description 14
- 239000011724 folic acid Substances 0.000 claims abstract description 14
- 201000003444 follicular lymphoma Diseases 0.000 claims abstract description 14
- 201000007924 marginal zone B-cell lymphoma Diseases 0.000 claims abstract description 14
- 208000021937 marginal zone lymphoma Diseases 0.000 claims abstract description 14
- 230000004108 pentose phosphate pathway Effects 0.000 claims abstract description 12
- FMGSKLZLMKYGDP-UHFFFAOYSA-N Dehydroepiandrosterone Natural products C1C(O)CCC2(C)C3CCC(C)(C(CC4)=O)C4C3CC=C21 FMGSKLZLMKYGDP-UHFFFAOYSA-N 0.000 claims description 43
- 229960002847 prasterone Drugs 0.000 claims description 43
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 claims description 24
- 150000003839 salts Chemical class 0.000 claims description 24
- IWUCUJZMKDHLOF-UHFFFAOYSA-N 6-amino-4-[3-(hydroxymethyl)-5-(5-hydroxypent-1-ynyl)phenyl]-3-methyl-4-propan-2-yl-2H-pyrano[2,3-c]pyrazole-5-carbonitrile Chemical group CC(C)C1(C2=C(NN=C2C)OC(N)=C1C#N)C1=CC(CO)=CC(=C1)C#CCCCO IWUCUJZMKDHLOF-UHFFFAOYSA-N 0.000 claims description 23
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 claims description 14
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 claims description 9
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 9
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 claims description 7
- 230000003042 antagnostic effect Effects 0.000 claims 1
- 229940116395 Serine hydroxymethyltransferase inhibitor Drugs 0.000 abstract description 7
- 210000004698 lymphocyte Anatomy 0.000 abstract description 5
- 230000001684 chronic effect Effects 0.000 abstract description 4
- 230000005764 inhibitory process Effects 0.000 abstract description 4
- 208000032839 leukemia Diseases 0.000 abstract description 4
- ACFIXJIJDZMPPO-NNYOXOHSSA-N NADPH Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](OP(O)(O)=O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 ACFIXJIJDZMPPO-NNYOXOHSSA-N 0.000 abstract 2
- 210000004027 cell Anatomy 0.000 description 76
- 210000003719 b-lymphocyte Anatomy 0.000 description 17
- 230000030833 cell death Effects 0.000 description 14
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 12
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 12
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 10
- 239000001963 growth medium Substances 0.000 description 10
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 9
- 239000003814 drug Substances 0.000 description 9
- 230000001225 therapeutic effect Effects 0.000 description 9
- 206010028980 Neoplasm Diseases 0.000 description 8
- 201000010099 disease Diseases 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 239000002904 solvent Substances 0.000 description 8
- 238000002560 therapeutic procedure Methods 0.000 description 8
- 229940079593 drug Drugs 0.000 description 7
- 239000004480 active ingredient Substances 0.000 description 6
- 239000003642 reactive oxygen metabolite Substances 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- 201000011510 cancer Diseases 0.000 description 5
- 230000001472 cytotoxic effect Effects 0.000 description 5
- 230000034994 death Effects 0.000 description 5
- 102100034606 Serine hydroxymethyltransferase, mitochondrial Human genes 0.000 description 4
- 230000006907 apoptotic process Effects 0.000 description 4
- 238000003149 assay kit Methods 0.000 description 4
- 231100000433 cytotoxic Toxicity 0.000 description 4
- 239000003937 drug carrier Substances 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000000861 pro-apoptotic effect Effects 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 230000009466 transformation Effects 0.000 description 4
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 108010090931 Proto-Oncogene Proteins c-bcl-2 Proteins 0.000 description 3
- 102000013535 Proto-Oncogene Proteins c-bcl-2 Human genes 0.000 description 3
- 101710197776 Serine hydroxymethyltransferase 2 Proteins 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 230000014509 gene expression Effects 0.000 description 3
- 230000013632 homeostatic process Effects 0.000 description 3
- 230000001613 neoplastic effect Effects 0.000 description 3
- 230000002018 overexpression Effects 0.000 description 3
- 238000011458 pharmacological treatment Methods 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 230000002195 synergetic effect Effects 0.000 description 3
- 206010067484 Adverse reaction Diseases 0.000 description 2
- 108010063104 Apoptosis Regulatory Proteins Proteins 0.000 description 2
- 102000010565 Apoptosis Regulatory Proteins Human genes 0.000 description 2
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 2
- 108091012583 BCL2 Proteins 0.000 description 2
- 101100016516 Caenorhabditis elegans hbl-1 gene Proteins 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 108091006146 Channels Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108010043428 Glycine hydroxymethyltransferase Proteins 0.000 description 2
- 206010066476 Haematological malignancy Diseases 0.000 description 2
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 2
- 230000006838 adverse reaction Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000000259 anti-tumor effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000003111 delayed effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 229960004679 doxorubicin Drugs 0.000 description 2
- 239000003862 glucocorticoid Substances 0.000 description 2
- 201000005787 hematologic cancer Diseases 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 230000037353 metabolic pathway Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- -1 phosphate pentose Chemical class 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 230000001235 sensitizing effect Effects 0.000 description 2
- 238000009097 single-agent therapy Methods 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 230000008685 targeting Effects 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- OGWKCGZFUXNPDA-XQKSVPLYSA-N vincristine Chemical compound C([N@]1C[C@@H](C[C@]2(C(=O)OC)C=3C(=CC4=C([C@]56[C@H]([C@@]([C@H](OC(C)=O)[C@]7(CC)C=CCN([C@H]67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)C[C@@](C1)(O)CC)CC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-XQKSVPLYSA-N 0.000 description 2
- 229960004528 vincristine Drugs 0.000 description 2
- OGWKCGZFUXNPDA-UHFFFAOYSA-N vincristine Natural products C1C(CC)(O)CC(CC2(C(=O)OC)C=3C(=CC4=C(C56C(C(C(OC(C)=O)C7(CC)C=CCN(C67)CC5)(O)C(=O)OC)N4C=O)C=3)OC)CN1CCC1=C2NC2=CC=CC=C12 OGWKCGZFUXNPDA-UHFFFAOYSA-N 0.000 description 2
- YPSXFMHXRZAGTG-UHFFFAOYSA-N 4-methoxy-2-[2-(5-methoxy-2-nitrosophenyl)ethyl]-1-nitrosobenzene Chemical compound COC1=CC=C(N=O)C(CCC=2C(=CC=C(OC)C=2)N=O)=C1 YPSXFMHXRZAGTG-UHFFFAOYSA-N 0.000 description 1
- 208000024893 Acute lymphoblastic leukemia Diseases 0.000 description 1
- 208000014697 Acute lymphocytic leukaemia Diseases 0.000 description 1
- 208000003950 B-cell lymphoma Diseases 0.000 description 1
- 239000012664 BCL-2-inhibitor Substances 0.000 description 1
- 241001103259 Baltimora Species 0.000 description 1
- 229940123711 Bcl2 inhibitor Drugs 0.000 description 1
- 102100023073 Calcium-activated potassium channel subunit alpha-1 Human genes 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 102100025064 Cellular tumor antigen p53 Human genes 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- 102000018832 Cytochromes Human genes 0.000 description 1
- 108010052832 Cytochromes Proteins 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 1
- 208000023661 Haematological disease Diseases 0.000 description 1
- 101001056180 Homo sapiens Induced myeloid leukemia cell differentiation protein Mcl-1 Proteins 0.000 description 1
- 101001122313 Homo sapiens Metalloendopeptidase OMA1, mitochondrial Proteins 0.000 description 1
- 101000686031 Homo sapiens Proto-oncogene tyrosine-protein kinase ROS Proteins 0.000 description 1
- 101000932478 Homo sapiens Receptor-type tyrosine-protein kinase FLT3 Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 1
- 102100026539 Induced myeloid leukemia cell differentiation protein Mcl-1 Human genes 0.000 description 1
- 239000002177 L01XE27 - Ibrutinib Substances 0.000 description 1
- 108010092555 Large-Conductance Calcium-Activated Potassium Channels Proteins 0.000 description 1
- 208000007466 Male Infertility Diseases 0.000 description 1
- 102100027104 Metalloendopeptidase OMA1, mitochondrial Human genes 0.000 description 1
- 150000001204 N-oxides Chemical class 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102100023347 Proto-oncogene tyrosine-protein kinase ROS Human genes 0.000 description 1
- 102100020718 Receptor-type tyrosine-protein kinase FLT3 Human genes 0.000 description 1
- 229940124639 Selective inhibitor Drugs 0.000 description 1
- 206010070834 Sensitisation Diseases 0.000 description 1
- 101710084636 Serine hydroxymethyltransferase Proteins 0.000 description 1
- 101710099809 Serine hydroxymethyltransferase, cytosolic Proteins 0.000 description 1
- 101710087362 Serine hydroxymethyltransferase, mitochondrial Proteins 0.000 description 1
- 102000019394 Serine hydroxymethyltransferases Human genes 0.000 description 1
- 208000029052 T-cell acute lymphoblastic leukemia Diseases 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- DTQVDTLACAAQTR-UHFFFAOYSA-M Trifluoroacetate Chemical compound [O-]C(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-M 0.000 description 1
- 108010078814 Tumor Suppressor Protein p53 Proteins 0.000 description 1
- 230000001594 aberrant effect Effects 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000002424 anti-apoptotic effect Effects 0.000 description 1
- 238000003782 apoptosis assay Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 108700000711 bcl-X Proteins 0.000 description 1
- 102000055104 bcl-X Human genes 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000008512 biological response Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 239000008199 coating composition Substances 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000000890 drug combination Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 238000013265 extended release Methods 0.000 description 1
- 206010016256 fatigue Diseases 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 230000004992 fission Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 210000004524 haematopoietic cell Anatomy 0.000 description 1
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-M hydrogensulfate Chemical compound OS([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-M 0.000 description 1
- XYFPWWZEPKGCCK-GOSISDBHSA-N ibrutinib Chemical compound C1=2C(N)=NC=NC=2N([C@H]2CN(CCC2)C(=O)C=C)N=C1C(C=C1)=CC=C1OC1=CC=CC=C1 XYFPWWZEPKGCCK-GOSISDBHSA-N 0.000 description 1
- 229960001507 ibrutinib Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000034727 intrinsic apoptotic signaling pathway Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000003563 lymphoid tissue Anatomy 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 210000001700 mitochondrial membrane Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000000066 myeloid cell Anatomy 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 230000006654 negative regulation of apoptotic process Effects 0.000 description 1
- 208000004235 neutropenia Diseases 0.000 description 1
- 231100001143 noxa Toxicity 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 229940124531 pharmaceutical excipient Drugs 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000004014 plasticizer Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229960004618 prednisone Drugs 0.000 description 1
- XOFYZVNMUHMLCC-ZPOLXVRWSA-N prednisone Chemical compound O=C1C=C[C@]2(C)[C@H]3C(=O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 XOFYZVNMUHMLCC-ZPOLXVRWSA-N 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000005522 programmed cell death Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000000541 pulsatile effect Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 229960004641 rituximab Drugs 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000008137 solubility enhancer Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000011255 standard chemotherapy Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229940037128 systemic glucocorticoids Drugs 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000011285 therapeutic regimen Methods 0.000 description 1
- 230000004797 therapeutic response Effects 0.000 description 1
- 206010043554 thrombocytopenia Diseases 0.000 description 1
- JOXIMZWYDAKGHI-UHFFFAOYSA-M toluene-4-sulfonate Chemical compound CC1=CC=C(S([O-])(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-M 0.000 description 1
- 231100000816 toxic dose Toxicity 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/415—1,2-Diazoles
- A61K31/4162—1,2-Diazoles condensed with heterocyclic ring systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/565—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol
- A61K31/566—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol having an oxo group in position 17, e.g. estrone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Definitions
- the present invention relates to a combination comprising the BH3-mimetic compound ABT-199 (venetoclax) and at least one inhibitor of NADPH production and/or pharmaceutically acceptable salts or derivatives thereof, pharmaceutical compositions comprising such combination and use of the combination of the invention and of compositions that contain it for the treatment of non-Hodgkin's B-cell lymphomas, preferably for the treatment of diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocyte leukaemia (CLL), marginal zone B-cell lymphoma or mantle cell lymphoma (MCL), and even more preferably for the treatment of DLBCL, also in the treatment of acute leukaemia, preferably in the treatment of acute myeloid leukaemia (AML).
- AML acute myeloid leukaemia
- ROS reactive oxygen species
- the members of the Bcl-2 protein family are the main mitochondrial-dependent apoptosis regulators in eukaryotic cells and this family is composed both of anti- apoptotic proteins (Bcl-XL, Bcl-2, Bcl-W, Bfl-1, Mcl-1) and pro-apoptotic proteins (Bak, Bax, Bid, Bim, Bad, Bik, Bmf, Noxa, Puma).
- the protein Bcl-2 covers different roles, both physiological and pathological; among the most significant include the inhibition of apoptosis which, if altered, can promote aberrant cell survival. Therefore, the inhibition of the protein Bcl-2 can reactivate cell death in pathological contexts, leading to a better therapeutic response in cancer patients.
- the involvement of the Bcl-2 proteins in a large number of solid and haematological malignancies has been described in the state of the art.
- Bcl- 2 which covers a central role in the survival of B lymphocytes, is expressed aberrantly in B-cell non-Hodgkin's lymphoma (tumours deriving from the transformation of B lymphocytes) (Leverson JD and Cojocari D (2016) Hematologic Tumor Cell Resistance to the BCL-2 Inhibitor Venetoclax: A Product of Its Microenvironment? Front. Oncol. 8:458. doi: 10.3389/ fonc.2018.00458).
- Bcl-2 covers an important role in the onset of chemoresistance in acute myeloid leukaemia (Metha SV, Shukla SN and Vora HH (2013) Overexpression of Bcl2 protein predicts chemoresistance in acute myeloid leukemia: its correlation with FLT3. Neoplasma. 60(6):666 -75. doi:10.4149/neo_2013_085).
- B-cell non-Hodgkin's lymphoma means haematological malignancies that primarily involve the lymphoid tissues. Based on the type, NHL can be characterised by an indolent progression and high aggressiveness.
- the term "B-cell non-Hodgkin's lymphoma” comprises numerous neoplastic diseases that derive from the transformation of B lymphocytes. The 5 most common types - in order of prevalence - are diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukaemia (CLL), marginal zone B-cell lymphoma and mantle cell lymphoma.
- AML Acute myeloid leukaemia
- AML represents the most common type of acute leukaemia in adults (about 80% of cases).
- AML can affect patients with pre-existing haemato logical disorders, as a result of therapies or, in most cases, de novo, with the abnormal proliferation of myeloid cells and the monoclonal evolution of the disease.
- progress in the treatment of AML has led to significant improvements in the treatment of younger patients, the prognosis in patients aged over 65, who represent most new cases, remains unfavourable (De Kouchkovsky, I. and Abdul-Hay, M. Acute myeloid leukemia: a comprehensive review and 2016 update. Blood Cancer J (2016) 6(7):e441. doi: 10.1038/bcj.2016.50).
- ABT-199 also known as venetoclax, whose formula is provided below, is a BH3-mimetic compound specific for Bcl-2 which, through the inhibition of this protein, promotes the opening of the channel formed by Bax/Bak in the outer mitochondrial membrane.
- the opening of this channel represents the critical event for the release of cytochrome C from the mitochondria, which is in turn necessary for the activation of the intrinsic apoptosis pathway (Pernas, L. & Scorrano, L. Mito-Morphosis: Mitochondrial Fusion, Fission, and Cristae Remodeling as Key Mediators of Cellular Function. Annu Rev Physiol (2016) vol. 78, pages 505-531, doi:10.1146/annurev-physiol-021115-105011).
- the overexpression of the anti-apoptotic protein Bcl-2 is a common strategy adopted by tumour cells to prevent apoptosis, thus causing resistance to anti-tumour therapy.
- ABT-199 has demonstrated high efficacy in the treatment of relapsed/ refractory chronic lymphocytic leukaemia (RR-CLL) with the deletion of the chromosome 17p and was approved by the US Food and Drug Administration (FDA) in 2016.
- RR-CLL chronic lymphocytic leukaemia
- FDA US Food and Drug Administration
- the efficacy of ABT-199 in other non-Hodgkin's lymphomas is very variable, with a response rate limited to 18% in diffuse large B-cell lymphoma (DLBCL) (Roberts AW, Huang D. Targeting BCL2 With BH3 Mimetics: Basic Science and Clinical Application of Venetoclax in Chronic Lymphocytic Leukemia and Related B Cell Malignancies. Clin Pharmacol Ther.
- DLBCL represents the most frequent type of B cell non-Hodgkin's lymphoma and is characterised by marked intrinsic resistance to ABT-199
- the treatment of patients affected by this disease is strongly limited in clinical practice with respect to neoplasms such as CLL, therefore the development of a new strategy for overcoming cancer cell resistance to anti-tumour therapy with ABT- 199 is extremely important.
- ABT-199 there is also a strongly felt need for a reduction in the doses of ABT-199 necessary for obtaining a therapeutic effect, in order to reduce the toxic effects of this active ingredient, which present in particular as neutropenia, nausea, anaemia, diarrhoea, upper airways infection, fatigue, thrombocytopenia, male infertility.
- the toxicity of ABT- 199 is reported also towards normal B lymphocytes, which represent the healthy counterpart of DLBCL. This characteristic of ABT-199 makes it necessary to have a strategy that can eliminate tumour cells with low drug doses, thus increasing the therapeutic window.
- An object of the present invention is to overcome the disadvantages of the prior art.
- an object of the present invention is to provide a new and inventive combination of active pharmaceutical ingredients for the treatment of B-cell non- Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukemia.
- BH3-mimetic compound ABT-199 venetoclax
- at least one inhibitor of NADPH production such as, for example, an inhibitor of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway, preferably dehydroepiandrosterone (DHEA, inhibitor of the first enzyme of the pentose phosphate pathway, PPP) and/ or pharmaceutically acceptable salts or derivatives thereof, SHIN2, a serine hydroxymethyltransferase inhibitor (enzyme that catalyses the entry of monocarbon units in the mitochondrial folate pathway) and/ or pharmaceutically acceptable salts or derivatives thereof, and through pharmaceutical compositions that comprise such combination together with one or more physiologically acceptable excipients.
- PPP pentose phosphate pathway
- DHEA dehydroepiandrosterone
- SHIN2 serine hydroxymethyltransferase inhibitor (enzyme that catalyses the entry of monocarbon units in the mitochondrial folate pathway) and/ or pharmaceutically acceptable salts
- the invention also relates to the combination comprising the BH3-mimetic compound ABT-199 (venetoclax) and at least one inhibitor of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway and/ or pharmaceutically acceptable salts or derivatives thereof that comprise such combination, for use in the treatment of B-cell non- Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukemia, according to the characteristics of the appended claims, which form an integral part of the present invention.
- Another aspect of the present invention proposes a combination comprising ABT-199 (venetoclax) and dehydroepiandrosterone (DHEA) and/or pharmaceutically acceptable salts or derivatives thereof and the pharmaceutical compositions that comprise such combination together with one or more physiologically acceptable excipients.
- ABT-199 venetoclax
- DHEA dehydroepiandrosterone
- This aspect also relates to the combination comprising ABT-199 (venetoclax) and dehydroepiandrosterone (DHEA) and/or pharmaceutically acceptable salts or derivatives thereof and the pharmaceutical compositions that comprise it, for use in the treatment of B-cell non-Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukemia.
- ABT-199 venetoclax
- DHEA dehydroepiandrosterone
- a further aspect of the present invention proposes a combination comprising ABT-199 (venetoclax) and SHIN2, a serine hydroxymethyltransferase inhibitor and/or pharmaceutically acceptable salts or derivatives thereof and the pharmaceutical compositions that comprise such combination together with one or more physiologically acceptable excipients.
- Said aspect also relates to the combination comprising ABT-199 (venetoclax) and SHIN2, a serine hydroxymethyltransferase inhibitor, and/or pharmaceutically acceptable salts or derivatives thereof and the pharmaceutical compositions that comprise it, for use in the treatment of B-cell non-Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukaemia.
- phosphate pentose pathway and “mitochondrial folate pathway” refer to two metabolic pathways that predominantly support the production of NADPH in the cells; pharmaceuticals that antagonise such metabolic pathways in tumour cells therefore act as NADPH production inhibitors.
- physiologically acceptable excipient relates to a substance without any pharmacological effect of its own and that does not produce adverse reactions when administered to a mammal, preferably to a human being.
- Physiologically acceptable excipients are well known in the art and are described, for example in the Handbook of Pharmaceutical Excipients, sixth-edition (2009), incorporated herein for reference purposes.
- pharmaceutically acceptable salts or derivatives relates to salts or derivatives that have biological efficacy and the properties of the salified or derivatised compound and that do not produce adverse reactions when administered to a mammal, preferably to a human being.
- Pharmaceutically acceptable salts may be inorganic or organic salts; examples of pharmaceutically acceptable salts include, but are not limited to: carbonate, hydrochloride, hydrobromide, sulfate, hydrogen sulfate, citrate, maleate, fumarate, trifluoro acetate, 2-naphtalene sulfonate and para-toluenesulfonate. Further information on pharmaceutically acceptable salts can be found in Handbook of pharmaceutical salts, P. Stahl, C. Wermuth, WILEY-VCH, 127-133, 2008, incorporated herein for reference purposes.
- Pharmaceutically acceptable derivatives include esters, ethers, and N-oxides.
- treat refers to a method for attenuating or abrogating a disease and/ or its associated symptoms, and to an administration method of pharmaceuticals with the aim of inducing a biological response.
- modulated relates to the ability of a compound to quantitatively or qualitatively modify a biological function such as, for example, the activity of the protein Bcl-2 or the production of NADPH and the homeostasis of ROSs.
- composition intends to comprise a product not comprising the ingredients specified in the specified quantities, and any product that results, directly or indirectly, from the combination of the ingredients specified in the specified quantities.
- “Pharmaceutically acceptable” means that the carrier, the diluent or the excipient must be compatible with the other components of the formulation and not harmful for the receiver.
- DLBCL is an acronym for "diffuse large B-cell lymphoma”.
- AML is an acronym for "acute myeloid leukemia”.
- ROS reactive oxygen species
- CLL is an acronym for “chronic lymphocytic leukemia”.
- DHEA is an acronym for “dehydroepiandrosterone”.
- SHMT is an acronym for "serine hydroxymethyltransferase”.
- SHIN2 is an inhibitor of the enzyme serine hydroxymethyltransferase.
- R-CHOP is an acronym in which "R” indicates the active ingredient rituximab; “C” is the active ingredient cyclophosphamide; “H” is the active ingredient “hydroxydaunorubicin”, also known as doxorubicin; “O” is the active ingredient Oncovin®, also known as vincristine; and “P” is the active ingredient prednisone.
- T-ALL is an acronym for "T-cell acute lymphoblastic leukemia”.
- Bcl-2 is an acronym for "B-cell lymphoma-2" which indicates a family of genes and the related proteins with pro- or anti-apoptotic activity.
- CC50 is a term that means “Cytotoxic Concentration 50” indicating the cytotoxic concentration of a compound that can cause the death of 50% of the cells constituting an in vitro cell population.
- CC30 is a term that means “Cytotoxic Concentration 30" indicating the cytotoxic concentration of a compound that can cause the death of 30% of the cells constituting an in vitro cell population.
- specific cell death means the percentage of cells that die following a specific pharmacological treatment.
- sensitised cell lines means cell lines that have become more susceptible to the cytotoxic action of ABT-199, thanks to combined treatment with at least one inhibitor of the pentose phosphate pathway (PPP) (e.g., dehydroepiandrotestore) or of the mitochondrial folate pathway (e.g., SHIN2).
- PPP pentose phosphate pathway
- SHIN2 mitochondrial folate pathway
- RR-CLL is an acronym that indicated refractory/ resistant chronic lymphatic leukaemia.
- B-cell non-Hodgkin's lymphoma means a series of neoplastic diseases that derive from the transformation of B lymphocytes.
- the 5 most common types of B-cell non-Hodgkin's lymphoma are diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukaemia (CLL), marginal zone B-cell lymphoma and mantle cell lymphoma.
- AML acute myeloid leukemia
- Figure 1 shows the values of CC50 obtained by treating 14 DLBCL cell lines and 6 AML cell lines with ABT-199 only.
- Figures 2A, 2B, 2C, 2D, 2E and 2F show the results obtained in terms of specific cell death (%), calculated through marking with propidium iodide and cytofluorometry, of DLBCL cell lines ( Figures 2A, 2B e 2C) and AML ( Figures 2D, 2E e 2F) treated with increasing concentrations of ABT-199 (1 nM - 10 mM) only (black circles) or in combination with DHEA at the concentration of 100 mM (white circles).
- Figure 2A shows 7 "sensitised” DLBCL cell lines
- Figure 2B shows 5 DLBCL lines sensitised to DHEA
- Figure 2C shows 2 DLBCL lines sensitive to DHEA
- Figure 2C shows 2 DLBCL lines sensitive to ABT-199
- Figure 2D shows 2 "sensitive-sensitised” AML cell lines
- Figure 2E shows 2 "resistant- sensitised” AML lines
- Figure 2F shows 2 "resistant-non sensitised” AML lines to ABT-199.
- Figure 3 shows the NADPH levels, expressed in picomoles (pmol), obtained through the use of the NADP/NADPH-GloTM Assay kit (Promega Corporation) in the "sensitisable” DLBCL cell line OCI Lyl9 and in the "resistant-sens disable” AML line KG-1 after treatment with the carrier (NT) only or with DHEA at the concentration of 100 mM.
- Figure 4A shows the combination index obtained in the "sensitisable" DLBCL line OCI Lyl9, treated with ABT-199 (10, 25, 50 and 100 nM) and DHEA (10, 25, 50 and 100 mM) separately and with the combination of ABT-199 and DHEA, maintaining a ratio between the concentrations of the two drugs equal to 1:1000.
- Figure 4B shows the combination index obtained in the "resistant-sensitisable" AML line KG-1, treated with ABT-199 (10, 25, 50 and 100 nM) and DHEA (10, 25, 50 and 100 mM) separately and with the combination of ABT-199 and DHEA, maintaining a ratio between the concentrations of the two drugs equal to 1:1000.
- Figure 5 shows the CC 30 values related to RR-CLL treated with ABT-199 only, B lymphocytes and "sensitised" DLBCL lines treated with ABT-199 only or in combination with DHEA at the concentration of 100 mM.
- the Figure also shows the therapeutic window related to ABT-199 in the RR-CLL (indicated with an asterisk) and the therapeutic window related to the combination of ABT-199 and DHEA in the "sensitisable" DLBCL (indicated with two asterisks).
- the CC 30 datum related to RR-CLL was extrapolated by Anderson, M. A. et al.
- the BCL2 selective inhibitor venetoclax induces rapid onset apoptosis of CLL cells in patients via a TP53 -independent mechanism.
- Figure 6A shows the results obtained in terms of specific cell death (%), calculated through marking with propidium iodide and cytofluorometry, in the "sensitisable" DLBCL cell line OCI Lyl9 treated with increasing concentrations of ABT-199 (1 nM - 10 mM) only (black circles) or in combination with SHIN2 at the concentration of 25 mM (white circles).
- Figure 6B shows the results obtained in terms of specific cell death (%), calculated through marking with propidium iodide and cytofluorometry, in the "resistant-sensitisable" AML cell line KG-1 treated with increasing concentrations of ABT-199 (1 nM - 10 mM) only (black circles) or in combination with SHIN2 at the concentration of 25 mM (white circles).
- Figure 7 shows the NADPH levels, expressed in picomoles (pmol), obtained through the use of the NADP/NADPH-GloTM Assay kit (Promega Corporation) in the "sensitisable” DLBCL cell line OCI Lyl9 and in the "resistant-sensitisable” AML cell line KG-1 after treatment with the carrier (NT) only or with SHIN2 at the concentration of 25 mM.
- Figure 8 shows the combination index obtained in the "sensitisable" DLBCL line OCI
- Lyl9 treated with ABT-199 (10, 25, 50 and 100 nM) and SHIN2 (2.5, 6.25, 12.5 and 25 mM) separately and with the combination of ABT-199 and DHEA, maintaining a ratio between the concentrations of the two drugs equal to 1:250.
- the present invention relates to a combination comprising the BH3-mimetic compound ABT-199 and at least one inhibitor of NADPH production such as, for example, an inhibitor of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway, and/or pharmaceutically acceptable salts or derivatives thereof and to the pharmaceutical compositions that comprise such combination; furthermore, said pharmaceutical combinations and compositions are the subject matter of the invention for use in the treatment of B-cell non-Hodgkin's lymphomas, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocyte leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, and acute leukaemia, preferably for the treatment of acute myeloid leukaemia.
- PPP pentose phosphate pathway
- Particularly preferred inhibitors of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway are dehydroepiandrosterone (DHEA) and/or pharmaceutically acceptable salts or derivatives thereof and SHIN2, a serine hydroxymethyltransferase inhibitor and/or pharmaceutically acceptable salts or derivatives thereof.
- DHEA dehydroepiandrosterone
- SHIN2 a serine hydroxymethyltransferase inhibitor and/or pharmaceutically acceptable salts or derivatives thereof.
- the invention relates to the combination comprising ABT-199 and dehydroepiandrosterone (DHEA) and/or pharmaceutically acceptable salts or derivatives thereof and to the pharmaceutical compositions that comprise it; furthermore, said combination and its pharmaceutical compositions are the subject matter of the invention for use in the treatment of B-cell non-Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukemia.
- DHEA dehydroepiandrosterone
- the invention relates to a combination comprising ABT-199 and dehydroepiandrosterone (DHEA) and/ or pharmaceutically acceptable salts or derivatives and to the pharmaceutical compositions that comprise it; furthermore, said combination and its pharmaceutical compositions are the subject matter of the invention for use in the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably for the treatment of acute myeloid leukaemia.
- DHEA dehydroepiandrosterone
- the invention relates to the combination comprising ABT- 199 and SHIN2, a serine hydroxymethyltransferase inhibitor and/ or pharmaceutically acceptable salts or derivatives thereof and to the pharmaceutical compositions that comprise it; furthermore, said combination and its pharmaceutical compositions are the subject matter of the invention for use in the treatment of B-cell non-Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukaemia.
- DHEA an inhibitor of the pentose phosphate pathway
- SHIN2 an inhibitor of the mitochondrial folate pathway
- compositions for use in the present invention in general comprise an effective quantity of the combination according to the invention and a suitable pharmaceutically acceptable carrier.
- the compositions can be prepared in a known way, which usually implies the mixing of the compounds of the combination of the invention with one or more pharmaceutically acceptable carriers and, if desired, in combination with other pharmaceutically active compounds, when necessary under aseptic conditions. Reference is made to the standard manuals, such as the latest edition of Remington's Pharmaceutical Sciences.
- the compounds may be formulated as a pharmaceutical preparation comprising at least one compound and at least one pharmaceutically acceptable carrier, diluent, or excipient and/ or adjuvant and optionally one or more further pharmaceutically active compounds.
- the pharmaceutical combinations of the present invention are preferably in the form of a single dose and can be appropriately packaged, e.g., in a box, blister pack, vial, bottle, sachet or any other suitable disposable or multi-dose medium or container (which can be correctly labelled); optionally with one or more leaflets containing information on the product and/ or instructions for use.
- Formulations containing the combination described herein can be prepared using a pharmaceutically acceptable carrier which is considered safe and effective and can be administered to an individual without causing undesirable biological effects or undesired interactions.
- the "carrier” comprises, but not only, diluents, binders, lubricants, disintegrators, fillers, pH modifying agents, preservatives, and solubility enhancers.
- the carrier also includes components of the coating composition which may include plasticizers, pigments, dyes, stabilising agents and glidants. Delayed release, extended release and/or pulsatile release dose formulations can be prepared as described in standard references such as "Compresse farmaceutiche per forme di dosaggio", ed. Liberman et al. (New York, Marcel Dekker, Inc., 1989), "Remington - La scienza e la pratica della farmacia", 20th edition (Lippincott Williams & Wilkins, Baltimora, Maryland, 2000) and "Forme di dosaggio farmaceutiche e isti di somministrazione di farmaci", 6th Edition, Ansel et al. (Media, PA: Williams e Wilkins, 1995). These references provide information on media, materials, equipment, and processes for the preparation of tablets and capsules and delayed release dose forms of tablets, capsules, and granules.
- a panel of 14 DLBCL cell lines were used, in particular the cell lines OCI Lyl, OCI Lyl9, SUDHL6, Karpas 422, Pfeiffer, OCI Ly7, SUDHL4, OCI LylO, U2932, RIVA, TMD8, OCI Ly3, HBL1 and U2946, and a panel of 6 AML cell lines, in particular the cell lines MOLM- 13, MV4-11, KG-1, OCI-AML3, HEL and MOLM-16.
- the CC50 values of the OCI Ly7, OCI-AML3, HEL and MOLM-16 cell lines are greater than 10 mM.
- DLBCL and AML cell lines were treated with increasing concentrations of ABT-199 (1, 10, 100, 1000 and 10000 nM) alone (black circles in Figures 2A, 2B, 2C, 2D, 2E and 2F) or in combination with DHEA at the concentration of 100 mM (white circles in Figures 2A, 2B, 2C, 2D, 2E and 2F).
- DHEA is dissolved in absolute ethanol at an initial concentration 1000 times higher than the final one.
- T indicates the percentage of cells treated with propidium iodide (PI)
- NT indicates the percentage of cells treated with the solvent positive to marking with PL
- the DLBCL cell lines can therefore be grouped into three categories according to their response to the combination of DHEA and ABT-199.
- the “sensitised” cell lines include OCI Lyl9, SUDHL6, Karpas 422, SUDHL4, U2932, OCI Ly3 and HBL1.
- the cell lines sensitive to DHEA include Pfeiffer, OCI Ly7, OCI LylO, TMD8 and U2946.
- the cell lines sensitive to ABT-199 include OCI Lyl and RIVA.
- AML cell lines can be grouped into three categories according to their response to the combination of DHEA and ABT-199.
- the "sensitive-sensitisable" cell lines include MOLM-13 and MV4-11.
- the "resistant-sensitisable" cell lines include KG-1 and OCI-AML3.
- resistant-non-sensitisable cell lines include HEL and MOLM-16.
- DHEA causes a drastic sensitisation to ABT- 199 of a subgroup of DLBCL cell lines (with an increase of CC50 of about 700 times, Figure 2A).
- DHEA was able to reinforce the pro-apoptotic effects of ABT-199 (Figure 2C), and even cell lines that are completely refractory to ABT-199 displayed high sensitivity to DHEA ( Figure 2B).
- the addition of DHEA sensitises a subgroup of AML cell lines to ABT-199 (with an increase of CC50 of about 95 times, Figure 2D).
- these figures show the combination index obtained in the "sensitisable" DLBCL cell line OCI Lyl9 and in the "resistant-sensitisable” AML line KG-1.
- the cells were seeded (lxl0 6 cells/ml of culture medium) in plates of culture medium with 48 wells and treated with concentrations of ABT-199 equal to 10, 25, 50 and 100 nM, of DHEA equal to 10, 25, 50 and 100 mM, and with a combination of ABT-199 and DHEA, maintaining a ratio between the concentrations of the two drugs equal to 1:1000.
- ABT-199 The various concentrations of ABT-199 were prepared in the solvent dimethylsulfoxide (DMSO) and those of DHEA in the solvent absolute ethanol, at an initial concentration 1000 times higher than the final one, so that in all cases the solvent is diluted 1:1000 in the culture medium.
- DMSO solvent dimethylsulfoxide
- Figure 5 shows the values of CC30 obtained from literature for RR-CLL and for experiments performed in the laboratory for healthy B lymphocytes and for "sensitised” DLBCL lines.
- the specific cell death (%) was measured by marking with propidium iodide and cytofluorometry (as indicated above) after treatment of the cells with increasing concentrations of ABT-199 (1, 10, 100, 1000 and 10000 nM) alone or in combination with DHEA at the concentration of 100 mM.
- the B lymphocytes used in these experiments were isolated starting from mononuclear lymphocytes deriving from peripheral blood of healthy donors through immunomagnetic selection using the "B Cell Isolation Kit II, Human” kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions.
- the therapeutic windows indicated in Figure 5 relate to the efficacy of ABT-199 in RR- CLL with respect to healthy B lymphocytes and to the efficacy of the combination of ABT- 199 and DHEA in "sensitised" DLBCL lines with respect to healthy B lymphocytes treated likewise.
- the resistance to ABT-199 of the DLBCL lines with respect to B lymphocytes can be deduced (CC30 about 4.7 times higher in DLBCL lines with respect to B lymphocytes).
- the CC30 value of the RR-CLL treated with ABT-199 is about 5.5 times lower with respect to the value observed in B lymphocytes (defining the therapeutic window of ABT-199 in the RR-CLL).
- CC30 of the "sensitised" lines of DLBCL treated with the combination of ABT-199 and DHEA is 932 times lower than the value observed in B lymphocytes (defining the therapeutic window of the combination of ABT-199 and DHEA in "sensitisable” DLBCL).
- T indicates the percentage of cells treated with propidium iodide (PI)
- NT indicates the percentage of cells treated with the solvent positive to marking with PL
- SHIN2 serine hydroxymethyltransferase 2
- SHIN2 is able to sensitise the DLBCL and AML cells to the death induced by ABT-199 ( Figure 6A and 6B) and to induce a drop in NADPH levels associated with the death observed ( Figure 7).
- the Figures show the specific cell death (%), measured in cytofluorometry through marking with propidium iodide in the "sensitisable" DLBCL cell line OCI Lyl9 ( Figure 6A) and in the "resistant-sens disable” AML line KG-1 ( Figure 6B), keeping the culture conditions unvaried, following treatment with increasing combinations of ABT-199 (1, 10, 100, 1000 and 10000 nM) alone (black circles) or in combination with SHIN2 at the concentration 25 mM (white circles).
- the specific cell death was calculated as ((T- NT)/(100-NT))*100, where T indicates the percentage of cells treated with propidium iodide (PI) and NT indicates the percentage of cells treated with the solvent positive to marking with PI.
- Figure 5B shows the levels of NADPH, expressed in picomoles (pmol), measured with the NADP/NADPH-GloTM Assay kit (Promega Corporation) in the "sensitised" DLBCL cell line OCI Lyl9 and in the "resistant-sensitisable" AML line KG- 1.
- the OCI Lyl9 e KG-1 cells seeded at a concentration equal to lxlO 6 cells/ml of culture medium with 6 wells, were treated with SHIN2 at the concentration of 25 mM.
- ABT-199 and SHIN2 were prepared in the carrier dimethylsulfoxide (DMSO) at an initial concentration 1000 times higher than the final one, so that in all cases the solvent is diluted 1:1000 in the culture medium. Therefore, it can be concluded that SHIN2 also acts in synergy with ABT-199, sensitising the DLBCL cell lines to the action of ABT-199.
- DMSO carrier dimethylsulfoxide
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Oncology (AREA)
- Hematology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention relates to a combination comprising the BH3-mimetic compound ABT-199 (venetoclax) and at least one inhibitor of NADPH production, pharmaceutical compositions comprising such combination and use in the treatment of non-Hodgkin's B-cell lymphomas, preferably for the treatment of diffuse large B- cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocyte leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, even more preferably for the treatment of DLBCL, also in the treatment of acute leukaemia, preferably in the treatment of acute myeloid leukaemia (AML). In particular, the inhibition of NADPH production can take place, for example, through an inhibitor of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway. More in particular, the invention relates to a combination comprising ABT-199 and dehydroepiandrosterone (DHEA) or serine hydroxymethyltransferase inhibitor (SHIN2) and pharmaceutical compositions that comprise it, for use in the treatment of non- Hodgkin's B-cell lymphomas, preferably for the treatment of diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocyte leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, even more preferably for the treatment of DLBCL, also in the treatment of acute leukaemia, preferably in the treatment of acute myeloid leukaemia (AML).
Description
COMPOSITION COMPRISING THE BH3-MIMETIC ABT-199 (VENETOCLAX) AND AT LEAST ONE INHIBITOR OF THE NADPH PRODUCTION
DESCRIPTION
TECHNICAL FIELD
The present invention relates to a combination comprising the BH3-mimetic compound ABT-199 (venetoclax) and at least one inhibitor of NADPH production and/or pharmaceutically acceptable salts or derivatives thereof, pharmaceutical compositions comprising such combination and use of the combination of the invention and of compositions that contain it for the treatment of non-Hodgkin's B-cell lymphomas, preferably for the treatment of diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocyte leukaemia (CLL), marginal zone B-cell lymphoma or mantle cell lymphoma (MCL), and even more preferably for the treatment of DLBCL, also in the treatment of acute leukaemia, preferably in the treatment of acute myeloid leukaemia (AML).
BACKGROUND
It has been described how most tumour cells have higher levels of reactive oxygen species (ROS) with respect to normal cells (Kumari, S., Badana, A. K., G, M. M., G, S. & Malla, R. in Biomark Insights (2018) Vol. 13). Tumour cells could therefore be vulnerable to a further increase in ROS that activates programmed cell death (apoptosis). Such increase in ROS levels can be obtained through pharmacological interventions that aim to reduce the levels of NADPH, the production of which is mainly supported by the pentose phosphate pathway (PPP) and by the mitochondrial folate pathway (Ciccarese, F., Ciminale, V. Escaping death: mitochondrial redox homeostasis in cancer cells. Front Oncol (2017) vol 7, p. 117, doi: 10.3389/fonc.2017.00117).
For example, it has been demonstrated that an activator of the BK channel of potassium (compound NS1619) and an inhibitor of the pentose phosphate pathway (PPP) (deh droepiandrosterone - DHEA) are able to sensitise T-cell acute lymphoblastic leukaemia cells (T-ALL) at cell death induced by glucocorticoids. This result is of particular interest as about 30% of patients with pediatric T-ALL is currently incurable, because of primary or secondary resistance to glucocorticoid based therapies (Silic- Benussi, M. et al. Selective killing of human T-ALL cells: an integrated approach targeting redox homeostasis and the OMA1/OPA1 axis. Cell Death Dis (2018) vol 9, p. 822, doi:10.1038/s41419-018-0870-9).
The members of the Bcl-2 protein family are the main mitochondrial-dependent apoptosis regulators in eukaryotic cells and this family is composed both of anti- apoptotic proteins (Bcl-XL, Bcl-2, Bcl-W, Bfl-1, Mcl-1) and pro-apoptotic proteins (Bak, Bax, Bid, Bim, Bad, Bik, Bmf, Noxa, Puma).
In general, the protein Bcl-2 covers different roles, both physiological and pathological; among the most significant include the inhibition of apoptosis which, if altered, can promote aberrant cell survival. Therefore, the inhibition of the protein Bcl-2 can reactivate cell death in pathological contexts, leading to a better therapeutic response in cancer patients. In fact, the involvement of the Bcl-2 proteins in a large number of solid and haematological malignancies has been described in the state of the art. In particular, Bcl- 2, which covers a central role in the survival of B lymphocytes, is expressed aberrantly in B-cell non-Hodgkin's lymphoma (tumours deriving from the transformation of B lymphocytes) (Leverson JD and Cojocari D (2018) Hematologic Tumor Cell Resistance to the BCL-2 Inhibitor Venetoclax: A Product of Its Microenvironment? Front. Oncol. 8:458. doi: 10.3389/ fonc.2018.00458). It is also known that the overexpression of Bcl-2 covers an important role in the onset of chemoresistance in acute myeloid leukaemia (Metha SV, Shukla SN and Vora HH (2013) Overexpression of Bcl2 protein predicts chemoresistance in acute myeloid leukemia: its correlation with FLT3. Neoplasma. 60(6):666 -75. doi:10.4149/neo_2013_085).
B-cell non-Hodgkin's lymphoma (NHL) means haematological malignancies that primarily involve the lymphoid tissues. Based on the type, NHL can be characterised by an indolent progression and high aggressiveness. The term "B-cell non-Hodgkin's lymphoma" comprises numerous neoplastic diseases that derive from the transformation of B lymphocytes. The 5 most common types - in order of prevalence - are diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukaemia (CLL), marginal zone B-cell lymphoma and mantle cell lymphoma.
Acute myeloid leukaemia (AML) represents the most common type of acute leukaemia in adults (about 80% of cases). AML can affect patients with pre-existing haemato logical disorders, as a result of therapies or, in most cases, de novo, with the abnormal proliferation of myeloid cells and the monoclonal evolution of the disease. Although progress in the treatment of AML has led to significant improvements in the treatment of younger patients, the prognosis in patients aged over 65, who represent most new cases, remains unfavourable (De Kouchkovsky, I. and Abdul-Hay, M. Acute myeloid leukemia: a comprehensive review and 2016 update. Blood Cancer J (2016) 6(7):e441. doi: 10.1038/bcj.2016.50).
ABT-199, also known as venetoclax, whose formula is provided below, is a BH3-mimetic compound specific for Bcl-2 which, through the inhibition of this protein, promotes the opening of the channel formed by Bax/Bak in the outer mitochondrial membrane. The opening of this channel represents the critical event for the release of cytochrome C from the mitochondria, which is in turn necessary for the activation of the intrinsic apoptosis pathway (Pernas, L. & Scorrano, L. Mito-Morphosis: Mitochondrial Fusion, Fission, and Cristae Remodeling as Key Mediators of Cellular Function. Annu Rev Physiol (2016) vol. 78, pages 505-531, doi:10.1146/annurev-physiol-021115-105011).
(venetoclax): nomenclature IUPAC 4-[4-[[2-(4-chlorophenyl)-4,4-dimethylcyclohexene-l- yl]methyl]piperazin-l-yl]-N-[3-nitro-4-(oxan-4-ylmethylamino)phenyl]sulfonyl-2-(lH- pyrrolo [2,3- b] pyridin-5-yloxy)benzamide.
The overexpression of the anti-apoptotic protein Bcl-2 is a common strategy adopted by tumour cells to prevent apoptosis, thus causing resistance to anti-tumour therapy.
The use of ABT-199 has demonstrated high efficacy in the treatment of relapsed/ refractory chronic lymphocytic leukaemia (RR-CLL) with the deletion of the
chromosome 17p and was approved by the US Food and Drug Administration (FDA) in 2016. Despite this, the efficacy of ABT-199 in other non-Hodgkin's lymphomas is very variable, with a response rate limited to 18% in diffuse large B-cell lymphoma (DLBCL) (Roberts AW, Huang D. Targeting BCL2 With BH3 Mimetics: Basic Science and Clinical Application of Venetoclax in Chronic Lymphocytic Leukemia and Related B Cell Malignancies. Clin Pharmacol Ther. 2017;101(l):89-98). As DLBCL represents the most frequent type of B cell non-Hodgkin's lymphoma and is characterised by marked intrinsic resistance to ABT-199, the treatment of patients affected by this disease is strongly limited in clinical practice with respect to neoplasms such as CLL, therefore the development of a new strategy for overcoming cancer cell resistance to anti-tumour therapy with ABT- 199 is extremely important.
There is also a strongly felt need for a reduction in the doses of ABT-199 necessary for obtaining a therapeutic effect, in order to reduce the toxic effects of this active ingredient, which present in particular as neutropenia, nausea, anaemia, diarrhoea, upper airways infection, fatigue, thrombocytopenia, male infertility. In particular, the toxicity of ABT- 199 is reported also towards normal B lymphocytes, which represent the healthy counterpart of DLBCL. This characteristic of ABT-199 makes it necessary to have a strategy that can eliminate tumour cells with low drug doses, thus increasing the therapeutic window.
For the treatment of DLBCL, therapeutic regimens have been proposed based on the combination between standard chemotherapy (R-CHOP) and ibrutinib which, although demonstrating significant progress, have not enabled the percentage to be decreased in about 40% of patients affected by DLBCL who are therefore currently incurable.
Therefore, considering that about 40% of patients affected by DLBCL do not respond to therapies currently available and considering the high intrinsic resistance of DLBCL to ABT-199, it is necessary to develop new therapies for patients with refractory DLBCL.
Furthermore, a phase 2 clinical trial demonstrated that ABT-199 in monotherapy leads to complete remission in only 19% of cases of acute myeloid leukemia (AML) already previously treated with other therapies (Konopleva, M. et al. Efficacy and biological correlates of response in a phase II study of venetoclax monotherapy in patients with acute myelogenous leukemia. Cancer Discov (2016) vol. 10, 1106- 1117, doi: 10.1158/2159- 8290. CD-16-0313), indicating the high resistance of this disease to treatment with ABT- 199.
Therefore, the development of new therapies is extremely important for the treatment of
patients affected by refractory AML.
OBJECTS AND SUMMARY OF THE INVENTION
An object of the present invention is to overcome the disadvantages of the prior art.
In particular, an object of the present invention is to provide a new and inventive combination of active pharmaceutical ingredients for the treatment of B-cell non- Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukemia.
These and other purposes of the present invention are reached by means of a combination comprising the BH3-mimetic compound ABT-199 (venetoclax) and at least one inhibitor of NADPH production such as, for example, an inhibitor of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway, preferably dehydroepiandrosterone (DHEA, inhibitor of the first enzyme of the pentose phosphate pathway, PPP) and/ or pharmaceutically acceptable salts or derivatives thereof, SHIN2, a serine hydroxymethyltransferase inhibitor (enzyme that catalyses the entry of monocarbon units in the mitochondrial folate pathway) and/ or pharmaceutically acceptable salts or derivatives thereof, and through pharmaceutical compositions that comprise such combination together with one or more physiologically acceptable excipients.
The invention also relates to the combination comprising the BH3-mimetic compound ABT-199 (venetoclax) and at least one inhibitor of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway and/ or pharmaceutically acceptable salts or derivatives thereof that comprise such combination, for use in the treatment of B-cell non- Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukemia, according to the characteristics of the appended claims, which form an integral part of the present invention.
Another aspect of the present invention proposes a combination comprising ABT-199 (venetoclax) and dehydroepiandrosterone (DHEA) and/or pharmaceutically acceptable salts or derivatives thereof and the pharmaceutical compositions that comprise such combination together with one or more physiologically acceptable excipients. This aspect
also relates to the combination comprising ABT-199 (venetoclax) and dehydroepiandrosterone (DHEA) and/or pharmaceutically acceptable salts or derivatives thereof and the pharmaceutical compositions that comprise it, for use in the treatment of B-cell non-Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukemia.
A further aspect of the present invention proposes a combination comprising ABT-199 (venetoclax) and SHIN2, a serine hydroxymethyltransferase inhibitor and/or pharmaceutically acceptable salts or derivatives thereof and the pharmaceutical compositions that comprise such combination together with one or more physiologically acceptable excipients.
Said aspect also relates to the combination comprising ABT-199 (venetoclax) and SHIN2, a serine hydroxymethyltransferase inhibitor, and/or pharmaceutically acceptable salts or derivatives thereof and the pharmaceutical compositions that comprise it, for use in the treatment of B-cell non-Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukaemia.
Further features and advantages of the present invention will be more apparent from the description of the accompanying drawings.
DEFINITIONS
Unless defined otherwise, all the terms of the art, notations and other scientific terms used herein are intended to have the meanings commonly understood by those who are skilled in the art to which this description pertains. In some cases, terms with meanings commonly understood are defined herein for clarity and/ or quick reference; the inclusion of such definitions in the present description need not therefore be interpreted as representative of a substantial difference with respect to what is generally understood in the art.
The terms "phosphate pentose pathway" and "mitochondrial folate pathway" refer to two metabolic pathways that predominantly support the production of NADPH in the cells; pharmaceuticals that antagonise such metabolic pathways in tumour cells therefore
act as NADPH production inhibitors.
The term "physiologically acceptable excipient" relates to a substance without any pharmacological effect of its own and that does not produce adverse reactions when administered to a mammal, preferably to a human being. Physiologically acceptable excipients are well known in the art and are described, for example in the Handbook of Pharmaceutical Excipients, sixth-edition (2009), incorporated herein for reference purposes.
The term "pharmacologically acceptable salts or derivatives" relates to salts or derivatives that have biological efficacy and the properties of the salified or derivatised compound and that do not produce adverse reactions when administered to a mammal, preferably to a human being. Pharmaceutically acceptable salts may be inorganic or organic salts; examples of pharmaceutically acceptable salts include, but are not limited to: carbonate, hydrochloride, hydrobromide, sulfate, hydrogen sulfate, citrate, maleate, fumarate, trifluoro acetate, 2-naphtalene sulfonate and para-toluenesulfonate. Further information on pharmaceutically acceptable salts can be found in Handbook of pharmaceutical salts, P. Stahl, C. Wermuth, WILEY-VCH, 127-133, 2008, incorporated herein for reference purposes. Pharmaceutically acceptable derivatives include esters, ethers, and N-oxides.
The terms "treat", "treating" and "treatment" refer to a method for attenuating or abrogating a disease and/ or its associated symptoms, and to an administration method of pharmaceuticals with the aim of inducing a biological response.
The term "modulated" relates to the ability of a compound to quantitatively or qualitatively modify a biological function such as, for example, the activity of the protein Bcl-2 or the production of NADPH and the homeostasis of ROSs.
The term "composition" as used in the present document intends to comprise a product not comprising the ingredients specified in the specified quantities, and any product that results, directly or indirectly, from the combination of the ingredients specified in the specified quantities. "Pharmaceutically acceptable" means that the carrier, the diluent or the excipient must be compatible with the other components of the formulation and not harmful for the receiver.
The term "simultaneous, separate or sequential use" relates to the simultaneous administration of the first and second compound or in such a way that the two compounds act in the patient's organism at the same time or to the administration of a compound after the other compound so as to provide a therapeutic effect.
The terms "comprising", "having", "including" and "containing" are to be considered as open terms (i.e., the meaning "comprising, but not limited to") and are to be considered as a support also for the terms "essentially consist of", "essentially consisting of", "consist of" or "consisting of".
"DLBCL" is an acronym for "diffuse large B-cell lymphoma".
"AML" is an acronym for "acute myeloid leukemia".
"ROS" is an acronym for "reactive oxygen species".
"CLL" is an acronym for "chronic lymphocytic leukemia".
"DHEA" is an acronym for "dehydroepiandrosterone".
"SHMT" is an acronym for "serine hydroxymethyltransferase".
"SHIN2" is an inhibitor of the enzyme serine hydroxymethyltransferase.
"R-CHOP" is an acronym in which "R" indicates the active ingredient rituximab; "C" is the active ingredient cyclophosphamide; "H" is the active ingredient "hydroxydaunorubicin", also known as doxorubicin; "O" is the active ingredient Oncovin®, also known as vincristine; and "P" is the active ingredient prednisone.
"T-ALL" is an acronym for "T-cell acute lymphoblastic leukemia".
"Bcl-2" is an acronym for "B-cell lymphoma-2" which indicates a family of genes and the related proteins with pro- or anti-apoptotic activity.
"CC50" is a term that means "Cytotoxic Concentration 50" indicating the cytotoxic concentration of a compound that can cause the death of 50% of the cells constituting an in vitro cell population.
"CC30" is a term that means "Cytotoxic Concentration 30" indicating the cytotoxic concentration of a compound that can cause the death of 30% of the cells constituting an in vitro cell population.
The expression "specific cell death" means the percentage of cells that die following a specific pharmacological treatment.
The expression "sensitised cell lines" means cell lines that have become more susceptible to the cytotoxic action of ABT-199, thanks to combined treatment with at least one
inhibitor of the pentose phosphate pathway (PPP) (e.g., dehydroepiandrotestore) or of the mitochondrial folate pathway (e.g., SHIN2).
The expression "therapeutic window" means the interval between the effective concentration and the toxic concentration of a pharmacological treatment. This parameter defines the safety of the pharmacological treatment itself in relation to the deriving clinical benefit. "RR-CLL" is an acronym that indicated refractory/ resistant chronic lymphatic leukaemia.
The term "B-cell non-Hodgkin's lymphoma" means a series of neoplastic diseases that derive from the transformation of B lymphocytes. The 5 most common types of B-cell non-Hodgkin's lymphoma are diffuse large B-cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukaemia (CLL), marginal zone B-cell lymphoma and mantle cell lymphoma.
The term "acute leukaemia" means a series of neoplastic diseases that originate from the transformation of haematopoietic cells of the lymphoid or myeloid line. The most common type of acute leukaemia in adults is represented by acute myeloid leukemia (AML).
BRIEF DESCRIPTION OF THE DRAWINGS
The invention will be described below with reference to some examples, provided by way of non-limiting example, and illustrated in the appended figures.
Figure 1 shows the values of CC50 obtained by treating 14 DLBCL cell lines and 6 AML cell lines with ABT-199 only.
Figures 2A, 2B, 2C, 2D, 2E and 2F show the results obtained in terms of specific cell death (%), calculated through marking with propidium iodide and cytofluorometry, of DLBCL cell lines (Figures 2A, 2B e 2C) and AML (Figures 2D, 2E e 2F) treated with increasing concentrations of ABT-199 (1 nM - 10 mM) only (black circles) or in combination with DHEA at the concentration of 100 mM (white circles). Figure 2A shows 7 "sensitised" DLBCL cell lines, Figure 2B shows 5 DLBCL lines sensitised to DHEA, Figure 2C shows 2 DLBCL lines sensitive to DHEA, Figure 2C shows 2 DLBCL lines sensitive to ABT-199, Figure 2D shows 2 "sensitive-sensitised" AML cell lines, Figure 2E shows 2 "resistant- sensitised" AML lines and Figure 2F shows 2 "resistant-non sensitised" AML lines to ABT-199.
Figure 3 shows the NADPH levels, expressed in picomoles (pmol), obtained through the use of the NADP/NADPH-Glo™ Assay kit (Promega Corporation) in the "sensitisable" DLBCL cell line OCI Lyl9 and in the "resistant-sens disable" AML line KG-1 after treatment with the carrier (NT) only or with DHEA at the concentration of 100 mM.
Figure 4A shows the combination index obtained in the "sensitisable" DLBCL line OCI Lyl9, treated with ABT-199 (10, 25, 50 and 100 nM) and DHEA (10, 25, 50 and 100 mM) separately and with the combination of ABT-199 and DHEA, maintaining a ratio between the concentrations of the two drugs equal to 1:1000. Figure 4B shows the combination index obtained in the "resistant-sensitisable" AML line KG-1, treated with ABT-199 (10, 25, 50 and 100 nM) and DHEA (10, 25, 50 and 100 mM) separately and with the combination of ABT-199 and DHEA, maintaining a ratio between the concentrations of the two drugs equal to 1:1000.
Figure 5 shows the CC30 values related to RR-CLL treated with ABT-199 only, B lymphocytes and "sensitised" DLBCL lines treated with ABT-199 only or in combination with DHEA at the concentration of 100 mM. The Figure also shows the therapeutic window related to ABT-199 in the RR-CLL (indicated with an asterisk) and the therapeutic window related to the combination of ABT-199 and DHEA in the "sensitisable" DLBCL (indicated with two asterisks). The CC30 datum related to RR-CLL was extrapolated by Anderson, M. A. et al. The BCL2 selective inhibitor venetoclax induces rapid onset apoptosis of CLL cells in patients via a TP53 -independent mechanism. Blood (2016) vol. 127, pages 3215-3224, doi:10.1182/blood-2016-01-688796.
Figure 6A shows the results obtained in terms of specific cell death (%), calculated through marking with propidium iodide and cytofluorometry, in the "sensitisable" DLBCL cell line OCI Lyl9 treated with increasing concentrations of ABT-199 (1 nM - 10 mM) only (black circles) or in combination with SHIN2 at the concentration of 25 mM (white circles). Figure 6B shows the results obtained in terms of specific cell death (%), calculated through marking with propidium iodide and cytofluorometry, in the "resistant-sensitisable" AML cell line KG-1 treated with increasing concentrations of ABT-199 (1 nM - 10 mM) only (black circles) or in combination with SHIN2 at the concentration of 25 mM (white circles).
Figure 7 shows the NADPH levels, expressed in picomoles (pmol), obtained through the use of the NADP/NADPH-Glo™ Assay kit (Promega Corporation) in the "sensitisable" DLBCL cell line OCI Lyl9 and in the "resistant-sensitisable" AML cell line KG-1 after treatment with the carrier (NT) only or with SHIN2 at the concentration of 25 mM.
Figure 8 shows the combination index obtained in the "sensitisable" DLBCL line OCI
Lyl9, treated with ABT-199 (10, 25, 50 and 100 nM) and SHIN2 (2.5, 6.25, 12.5 and 25 mM) separately and with the combination of ABT-199 and DHEA, maintaining a ratio between the concentrations of the two drugs equal to 1:250.
DETAILED DESCRIPTION OF THE INVENTION
The present invention relates to a combination comprising the BH3-mimetic compound ABT-199 and at least one inhibitor of NADPH production such as, for example, an inhibitor of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway, and/or pharmaceutically acceptable salts or derivatives thereof and to the pharmaceutical compositions that comprise such combination; furthermore, said pharmaceutical combinations and compositions are the subject matter of the invention for use in the treatment of B-cell non-Hodgkin's lymphomas, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocyte leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, and acute leukaemia, preferably for the treatment of acute myeloid leukaemia.
Particularly preferred inhibitors of the pentose phosphate pathway (PPP) or of the mitochondrial folate pathway are dehydroepiandrosterone (DHEA) and/or pharmaceutically acceptable salts or derivatives thereof and SHIN2, a serine hydroxymethyltransferase inhibitor and/or pharmaceutically acceptable salts or derivatives thereof.
Therefore, in particular, the invention relates to the combination comprising ABT-199 and dehydroepiandrosterone (DHEA) and/or pharmaceutically acceptable salts or derivatives thereof and to the pharmaceutical compositions that comprise it; furthermore, said combination and its pharmaceutical compositions are the subject matter of the invention for use in the treatment of B-cell non-Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukemia.
According to a particularly preferred aspect, the invention relates to a combination comprising ABT-199 and dehydroepiandrosterone (DHEA) and/ or pharmaceutically acceptable salts or derivatives and to the pharmaceutical compositions that comprise it; furthermore, said combination and its pharmaceutical compositions are the subject
matter of the invention for use in the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably for the treatment of acute myeloid leukaemia.
According to another aspect, the invention relates to the combination comprising ABT- 199 and SHIN2, a serine hydroxymethyltransferase inhibitor and/ or pharmaceutically acceptable salts or derivatives thereof and to the pharmaceutical compositions that comprise it; furthermore, said combination and its pharmaceutical compositions are the subject matter of the invention for use in the treatment of B-cell non-Hodgkin's lymphoma, preferably for the treatment of diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma, and even more preferably for the treatment of diffuse large B-cell lymphoma, acute leukaemia, preferably in the treatment of acute myeloid leukaemia.
In fact, it has been surprisingly found that it is possible to sensitise DLBCL and AML cells to the pro-apoptotic action of ABT-199 using an inhibitor of NADPH production, i.e., DHEA ( an inhibitor of the pentose phosphate pathway) or SHIN2 (an inhibitor of the mitochondrial folate pathway).
Thus, it was possible to overcome the resistance of DLBCL and AML cell lines to ABT- 199, as demonstrated in the examples provided below.
The pharmaceutical compositions for use in the present invention in general comprise an effective quantity of the combination according to the invention and a suitable pharmaceutically acceptable carrier. The compositions can be prepared in a known way, which usually implies the mixing of the compounds of the combination of the invention with one or more pharmaceutically acceptable carriers and, if desired, in combination with other pharmaceutically active compounds, when necessary under aseptic conditions. Reference is made to the standard manuals, such as the latest edition of Remington's Pharmaceutical Sciences.
Generally, for pharmaceutical use, the compounds may be formulated as a pharmaceutical preparation comprising at least one compound and at least one pharmaceutically acceptable carrier, diluent, or excipient and/ or adjuvant and optionally one or more further pharmaceutically active compounds.
The pharmaceutical combinations of the present invention are preferably in the form of a single dose and can be appropriately packaged, e.g., in a box, blister pack, vial, bottle, sachet or any other suitable disposable or multi-dose medium or container (which can be correctly labelled); optionally with one or more leaflets containing information on the product and/ or instructions for use.
Formulations containing the combination described herein can be prepared using a pharmaceutically acceptable carrier which is considered safe and effective and can be administered to an individual without causing undesirable biological effects or undesired interactions. As generally used in the present document, the "carrier" comprises, but not only, diluents, binders, lubricants, disintegrators, fillers, pH modifying agents, preservatives, and solubility enhancers.
The carrier also includes components of the coating composition which may include plasticizers, pigments, dyes, stabilising agents and glidants. Delayed release, extended release and/or pulsatile release dose formulations can be prepared as described in standard references such as "Compresse farmaceutiche per forme di dosaggio", ed. Liberman et al. (New York, Marcel Dekker, Inc., 1989), "Remington - La scienza e la pratica della farmacia", 20th edition (Lippincott Williams & Wilkins, Baltimora, Maryland, 2000) and "Forme di dosaggio farmaceutiche e sistemi di somministrazione di farmaci", 6th Edition, Ansel et al. (Media, PA: Williams e Wilkins, 1995). These references provide information on media, materials, equipment, and processes for the preparation of tablets and capsules and delayed release dose forms of tablets, capsules, and granules.
The invention is illustrated below through experimental examples, which are not considered limiting for the purpose of the invention.
EXAMPLES
EXAMPLE 1
Demonstration of overcoming resistance to ABT-199 in DLBCL and AML cell lines using the combination of ABT-199 and DHEA.
A panel of 14 DLBCL cell lines were used, in particular the cell lines OCI Lyl, OCI Lyl9, SUDHL6, Karpas 422, Pfeiffer, OCI Ly7, SUDHL4, OCI LylO, U2932, RIVA, TMD8, OCI Ly3, HBL1 and U2946, and a panel of 6 AML cell lines, in particular the cell lines MOLM- 13, MV4-11, KG-1, OCI-AML3, HEL and MOLM-16.
These cell lines were seeded (lxlO6 cells/ ml of culture medium) in cell culture plates with 48 wells and treated with concentrations of ABT-199 equal to 1, 10, 100, 1000 e 10000 nM, obtaining the CC50 values shown in Figure 1. The various concentrations of ABT-199 were prepared in the carrier dimethylsulfoxide (DMSO) at an initial concentration 1000 times higher than the final one, so that in all cases the solvent is diluted 1:1000 in the culture medium. The broken line indicates the average value of CC50 (1,9 nM) shown for the RR- CLL (see Anderson, M. A. et al. mentioned above).
In compliance with the clinical characteristics of the respective diseases, these lines have demonstrated high resistance to ABT-199, with a mean CC50 of 2.2 mM in the DLBCL cell line and a mean CC50 of 4.4 mM in AML cell lines.
The CC50 values of the OCI Ly7, OCI-AML3, HEL and MOLM-16 cell lines are greater than 10 mM.
Then (keeping the seeding and cell culture conditions unvaried) the same DLBCL and AML cell lines were treated with increasing concentrations of ABT-199 (1, 10, 100, 1000 and 10000 nM) alone (black circles in Figures 2A, 2B, 2C, 2D, 2E and 2F) or in combination with DHEA at the concentration of 100 mM (white circles in Figures 2A, 2B, 2C, 2D, 2E and 2F). DHEA is dissolved in absolute ethanol at an initial concentration 1000 times higher than the final one.
The specific cell death was calculated as ((T-NT)/(100-NT))*100, where T indicates the percentage of cells treated with propidium iodide (PI) and NT indicates the percentage of cells treated with the solvent positive to marking with PL
The DLBCL cell lines can therefore be grouped into three categories according to their response to the combination of DHEA and ABT-199.
The "sensitised" cell lines include OCI Lyl9, SUDHL6, Karpas 422, SUDHL4, U2932, OCI Ly3 and HBL1.
The cell lines sensitive to DHEA include Pfeiffer, OCI Ly7, OCI LylO, TMD8 and U2946. The cell lines sensitive to ABT-199 include OCI Lyl and RIVA.
Furthermore, the AML cell lines can be grouped into three categories according to their response to the combination of DHEA and ABT-199.
The "sensitive-sensitisable" cell lines include MOLM-13 and MV4-11.
The "resistant-sensitisable" cell lines include KG-1 and OCI-AML3.
The "resistant-non-sensitisable" cell lines include HEL and MOLM-16.
The values of CC50 shown in Figures 2A, 2B, 2C, 2D, 2E and 2F are calculated on the mean of three independent experiments conducted in triplicate for each cell line.
It is interesting to note how the addition of DHEA causes a drastic sensitisation to ABT- 199 of a subgroup of DLBCL cell lines (with an increase of CC50 of about 700 times, Figure
2A). In the cell lines that are relatively sensitive to ABT-199, DHEA was able to reinforce the pro-apoptotic effects of ABT-199 (Figure 2C), and even cell lines that are completely refractory to ABT-199 displayed high sensitivity to DHEA (Figure 2B). Furthermore, the addition of DHEA sensitises a subgroup of AML cell lines to ABT-199 (with an increase of CC50 of about 95 times, Figure 2D). Also, in a subgroup of AML cell lines resistant to ABT-199, DHEA was able to reinforce the pro-apoptotic effects of ABT-199 (Figure 2E), and even the AML cell lines that were completely refractory to ABT-199 (defined as "resistant-non sensitising") displayed moderate sensitivity to DHEA (Figure 2F).
The measurement of the NADPH levels, expressed in picomoles (pmol), through the NADP/NADPH-Glo™ Assay kit (Promega Corporation) in the "sensitisable" DLBCL cell line OCI Lyl9 and in the "resistant-sensitisable" AML line, KG-1 demonstrated how DHEA induces in these cells a drop in the production of NADPH (Figure 3). The OCI Lyl9 and KG-1 cells, seeded at a concentration equal to lxlO6 cells/ml of culture medium with 6 wells, were treated with DHEA at the concentration of 100 mM.
The isobolographic analysis performed using the Chou and Talalay equation (Chou, T. C. Drug combination studies and their synergy quantification using the Chou-Talalay method. Cancer Res (2010) vol. 70, pages 440-446, doi:10.1158/0008-5472.can-09-1947) revealed how the effects provided by ABT-199 and DHEA on the cell death of DLBCL and of AML are synergistic (Figures 4A and 4B).
In fact, these figures show the combination index obtained in the "sensitisable" DLBCL cell line OCI Lyl9 and in the "resistant-sensitisable" AML line KG-1. The cells were seeded (lxl06cells/ml of culture medium) in plates of culture medium with 48 wells and treated with concentrations of ABT-199 equal to 10, 25, 50 and 100 nM, of DHEA equal to 10, 25, 50 and 100 mM, and with a combination of ABT-199 and DHEA, maintaining a ratio between the concentrations of the two drugs equal to 1:1000. The various concentrations of ABT-199 were prepared in the solvent dimethylsulfoxide (DMSO) and those of DHEA in the solvent absolute ethanol, at an initial concentration 1000 times higher than the final one, so that in all cases the solvent is diluted 1:1000 in the culture medium.
Despite the healthy B lymphocytes being relatively sensitive to ABT-199, treatment with DHEA was not shown to be toxic for these cells, a datum that can be deduced from Figure
5.
In fact, Figure 5 shows the values of CC30 obtained from literature for RR-CLL and for experiments performed in the laboratory for healthy B lymphocytes and for "sensitised"
DLBCL lines. The specific cell death (%) was measured by marking with propidium iodide and cytofluorometry (as indicated above) after treatment of the cells with increasing concentrations of ABT-199 (1, 10, 100, 1000 and 10000 nM) alone or in combination with DHEA at the concentration of 100 mM. The B lymphocytes used in these experiments were isolated starting from mononuclear lymphocytes deriving from peripheral blood of healthy donors through immunomagnetic selection using the "B Cell Isolation Kit II, Human" kit (Miltenyi Biotec, Bergisch Gladbach, Germany) according to the manufacturer's instructions.
The therapeutic windows indicated in Figure 5 relate to the efficacy of ABT-199 in RR- CLL with respect to healthy B lymphocytes and to the efficacy of the combination of ABT- 199 and DHEA in "sensitised" DLBCL lines with respect to healthy B lymphocytes treated likewise. From the Figure, the resistance to ABT-199 of the DLBCL lines with respect to B lymphocytes can be deduced (CC30 about 4.7 times higher in DLBCL lines with respect to B lymphocytes). The CC30 value of the RR-CLL treated with ABT-199 is about 5.5 times lower with respect to the value observed in B lymphocytes (defining the therapeutic window of ABT-199 in the RR-CLL). The value of CC30 of the "sensitised" lines of DLBCL treated with the combination of ABT-199 and DHEA is 932 times lower than the value observed in B lymphocytes (defining the therapeutic window of the combination of ABT-199 and DHEA in "sensitisable" DLBCL).
The specific cell death was calculated as ((T-NT)/(100-NT))*100, where T indicates the percentage of cells treated with propidium iodide (PI) and NT indicates the percentage of cells treated with the solvent positive to marking with PL
EXAMPLE 2
Demonstration of overcoming resistance to ABT-199 in DLBCL and AML cell lines using the combination of ABT-199 and SHIN2.
A different inhibitor of the production of NADPH was tested, SHIN2, having the enzyme SHMT2 (serine hydroxymethyltransferase 2) as the target. The SHMT2 catalyses the entry of monocarbon units into the mitochondrial folate pathway that supports the production of NADPH.
Like DHEA, SHIN2 is able to sensitise the DLBCL and AML cells to the death induced by ABT-199 (Figure 6A and 6B) and to induce a drop in NADPH levels associated with the death observed (Figure 7).
The Figures show the specific cell death (%), measured in cytofluorometry through marking with propidium iodide in the "sensitisable" DLBCL cell line OCI Lyl9 (Figure
6A) and in the "resistant-sens disable" AML line KG-1 (Figure 6B), keeping the culture conditions unvaried, following treatment with increasing combinations of ABT-199 (1, 10, 100, 1000 and 10000 nM) alone (black circles) or in combination with SHIN2 at the concentration 25 mM (white circles). The specific cell death was calculated as ((T- NT)/(100-NT))*100, where T indicates the percentage of cells treated with propidium iodide (PI) and NT indicates the percentage of cells treated with the solvent positive to marking with PI. Figure 5B shows the levels of NADPH, expressed in picomoles (pmol), measured with the NADP/NADPH-Glo™ Assay kit (Promega Corporation) in the "sensitised" DLBCL cell line OCI Lyl9 and in the "resistant-sensitisable" AML line KG- 1. The OCI Lyl9 e KG-1 cells, seeded at a concentration equal to lxlO6 cells/ml of culture medium with 6 wells, were treated with SHIN2 at the concentration of 25 mM.
The isobolographic analysis performed using the Chou and Talalay equation (mentioned above) revealed how the effects provided by ABT-199 and SHIN2 on cell death of DLBCL are synergistic (Figure 8). In fact, the Figure shows the combination index in the "sensitisable" DLBCL cell line OCI Lyl9 (Figure 8). The cells, seeded at a concentration equal to lxl06cells/ml of culture medium in plates of culture medium with 48 wells and treated with concentrations of ABT-199 equal to 10, 25, 50 and 100 nM, of SHIN2 equal to 2.5; 6.25; 12.5 and 25 mM, and with a combination of ABT-199 and SHIN2, maintaining a ratio between the concentrations of the two drugs equal to 1:250. The various concentrations of ABT-199 and SHIN2 were prepared in the carrier dimethylsulfoxide (DMSO) at an initial concentration 1000 times higher than the final one, so that in all cases the solvent is diluted 1:1000 in the culture medium. Therefore, it can be concluded that SHIN2 also acts in synergy with ABT-199, sensitising the DLBCL cell lines to the action of ABT-199.
These data demonstrate how the reduction of NADPH levels following the inhibition of the mitochondrial folate pathway has a synergistic effect in combination with ABT-199, like what is observed with DHEA.
Claims
1. A combination comprising the BH3-mimetic compound ABT-199 (venetoclax) and at least one inhibitor of the NADPH production and/ or pharmaceutically acceptable salts or derivatives thereof.
2. The combination according to claim 1, wherein the inhibitor of the NADPH production acts with antagonistic activity on the pentose phosphate pathway or the mitochondrial folate pathway.
3. The combination according to claim 2, wherein the at least one inhibitor of the pentose phosphate pathway is dehydroepiandrosterone and/or pharmaceutically acceptable salts or derivatives thereof.
4. The combination according to claim 2, wherein the at least one inhibitor of the mitochondrial folate pathway is SHIN2 and/ or pharmaceutically acceptable salts or derivatives thereof.
5. The pharmaceutical formulation comprising any one of the combinations according to claims 1 to 4 and one or more physiologically acceptable excipients.
6. The combination according to any one of claims 1 to 4, for use in the treatment of B-cell non-Hodgkin's lymphoma.
7. The combination according to any one of claims 1 to 4, for use in the treatment of acute leukaemia.
8. The combination according to claim 6, wherein the non-Hodgkin's lymphomas are selected from among diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma.
9. The combination according to claim 8, wherein the non-Hodgkin's lymphoma is diffuse large B-cell lymphoma.
10. The combination according to claim 7, wherein the acute leukaemia is acute myeloid leukaemia.
11. A pharmaceutical formulation according to claim 5, for use in the treatment of B- cell non-Hodgkin's lymphoma.
12. The pharmaceutical formulation according to claim 11, wherein the non- Hodgkin's lymphomas are selected from diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic leukaemia, marginal zone B-cell lymphoma or mantle cell lymphoma.
13. The pharmaceutical formulation according to claim 12, wherein the non- Hodgkin's lymphoma is diffuse large B-cell lymphoma.
14. The pharmaceutical formulation according to claim 5, for use in the treatment of acute leukaemia.
15. The pharmaceutical formulation according to claim 14, wherein the acute leukaemia is acute myeloid leukemia.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IT102019000023895A IT201900023895A1 (en) | 2019-12-13 | 2019-12-13 | COMBINATION INCLUDING THE BH3-MIMETIC COMPOUND ABT-199 (VENETOCLAX) AND AT LEAST ONE INHIBITOR OF THE PRODUCTION OF NADPH AND / OR ITS SALTS OR PHARMACEUTICALLY ACCEPTABLE DERIVATIVES, PHARMACEUTICAL COMPOSITIONS THAT INCLUDE THIS COMBINATION OF NADPH AND / OR ITS SALTS OR PHARMACEUTICALLY ACCEPTABLE DERIVATIVES, PHARMACEUTICAL COMPOSITIONS INCLUDING THIS COMBINATION OF NADPH-LINKU, PREFERABLY IN THE TREATMENT OF DIFFUSED LARGE B-CELL LYMPHOMA (DLBCL) |
| IT102019000023895 | 2019-12-13 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO2021116929A1 true WO2021116929A1 (en) | 2021-06-17 |
Family
ID=70155025
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/IB2020/061684 WO2021116929A1 (en) | 2019-12-13 | 2020-12-09 | Composition comprising the bh3-mimetic abt-199 (venetoclax) and at least one inhibitor of the nadph production |
Country Status (2)
| Country | Link |
|---|---|
| IT (1) | IT201900023895A1 (en) |
| WO (1) | WO2021116929A1 (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022028353A1 (en) * | 2020-08-06 | 2022-02-10 | 北京诺诚健华医药科技有限公司 | Heterocyclic compound as bcl-2 inhibitor |
| CN114099464A (en) * | 2021-11-04 | 2022-03-01 | 国科温州研究院(温州生物材料与工程研究所) | Targeting nanoparticles for treating leukemia and preparation method and application thereof |
| CN114949230A (en) * | 2022-06-13 | 2022-08-30 | 厦门大学附属第一医院 | Combined pharmaceutical composition for preventing and/or treating acute myeloid leukemia and application thereof |
| CN116240283A (en) * | 2022-09-27 | 2023-06-09 | 广州市妇女儿童医疗中心 | Application of OMA1 in reversing acute lymphoblastic leukemia drug resistance |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015095834A2 (en) * | 2013-12-20 | 2015-06-25 | Biomed Valley Discoveries, Inc. | Cancer treatments using erk1/2 and bcl-2 family inhibitors |
-
2019
- 2019-12-13 IT IT102019000023895A patent/IT201900023895A1/en unknown
-
2020
- 2020-12-09 WO PCT/IB2020/061684 patent/WO2021116929A1/en active Application Filing
Non-Patent Citations (2)
| Title |
|---|
| DUCKER GREGORY S. ET AL: "Human SHMT inhibitors reveal defective glycine import as a targetable metabolic vulnerability of diffuse large B-cell lymphoma", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES, vol. 114, no. 43, 9 October 2017 (2017-10-09), US, pages 11404 - 11409, XP055780679, ISSN: 0027-8424, DOI: 10.1073/pnas.1706617114 * |
| SAMUEL Y NG ET AL: "Selective Bcl-2 inhibition to treat chronic lymphocytic leukemia and non-Hodgkin lymphoma", CLINICAL ADVANCES IN HEMATOLOGY & ONCOLOGY, vol. 12, no. 4, 1 April 2014 (2014-04-01), US, pages 224, XP055263314, ISSN: 1543-0790 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022028353A1 (en) * | 2020-08-06 | 2022-02-10 | 北京诺诚健华医药科技有限公司 | Heterocyclic compound as bcl-2 inhibitor |
| CN114099464A (en) * | 2021-11-04 | 2022-03-01 | 国科温州研究院(温州生物材料与工程研究所) | Targeting nanoparticles for treating leukemia and preparation method and application thereof |
| CN114099464B (en) * | 2021-11-04 | 2023-09-08 | 国科温州研究院(温州生物材料与工程研究所) | Targeted nanoparticle for treating leukemia as well as preparation method and application thereof |
| CN114949230A (en) * | 2022-06-13 | 2022-08-30 | 厦门大学附属第一医院 | Combined pharmaceutical composition for preventing and/or treating acute myeloid leukemia and application thereof |
| CN116240283A (en) * | 2022-09-27 | 2023-06-09 | 广州市妇女儿童医疗中心 | Application of OMA1 in reversing acute lymphoblastic leukemia drug resistance |
| CN116240283B (en) * | 2022-09-27 | 2024-05-28 | 广州市妇女儿童医疗中心 | Application of OMA1 in reversing acute lymphoblastic leukemia drug resistance |
Also Published As
| Publication number | Publication date |
|---|---|
| IT201900023895A1 (en) | 2021-06-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2021116929A1 (en) | Composition comprising the bh3-mimetic abt-199 (venetoclax) and at least one inhibitor of the nadph production | |
| US20200368235A1 (en) | Ibrutinib combination therapy | |
| WO2010033771A2 (en) | Modulators of hsp70/dnak function and methods of use thereof | |
| RU2731908C2 (en) | Apilimod composition and methods of using it in treating melanoma | |
| US20170071903A1 (en) | Use of eribulin and mtor inhibitors as combination therapy for the treatment of cancer | |
| US20160120871A1 (en) | Pharmaceutical combinations of a pi3k inhibitor and a microtubule destabilizing agent | |
| BR112020011167A2 (en) | composition and method for treating peripheral t-cell lymphoma and cutaneous t-cell lymphoma | |
| CN114096254A (en) | Cardionium salts as cognitive function enhancers | |
| US20230060792A1 (en) | Targeting Mcl-1 to Enhance DNA Replication Stress Sensitivity for Cancer Therapy | |
| CN113329749A (en) | Combination therapy for the treatment of uveal melanoma | |
| WO2021152467A1 (en) | Combinations comprising bh3-mimetic compounds and inhibitors of early stages of autophagy for use in the treatment of non-hodgkin's b-cell lymphomas and lymphoid and/or myeloid leukemias | |
| EP3782620B1 (en) | Pharmaceutical composition comprising 1,2-naphthoquinone derivative for use in preventing or treating acute myeloid or lymphoblastic leukemia | |
| CA2557857C (en) | Cancer treatment with topoisomerase-ii inhibitor, a bis-dioxypiperazine and radiation | |
| NO320019B1 (en) | Oral gelatin capsule formulation of 1,2,4-benzotriazine oxides and use the gelatin capsules to prepare a medicament. | |
| JP2014513698A (en) | Use of arsenic for cancer treatment protection | |
| EP4349340A1 (en) | Use of pharmaceutical composition for treating lung cancer | |
| US20230061353A1 (en) | Abt-751 and ionizing radiation | |
| WO2022199656A1 (en) | Pharmaceutical combination, kit containing same, and use thereof | |
| US20230390278A1 (en) | Synergistic combinations of valprooic acid and quinacrine | |
| WO2024176131A1 (en) | Tead- and kras g12d-inhibitor combinations for treating cancer | |
| Chauhan et al. | Oxidative Stress in Cancer: Therapeutic Implications of Small-Molecule Kinase Inhibitors | |
| WO2023070199A1 (en) | Use of sodium trans-[tetrachloridobis(1h-indazole)ruthenate(iii)] to ameliorate proteasome inhibitor resistance | |
| WO2024176130A1 (en) | Tead- and her2-inhibitor combinations for treating cancer | |
| JPH0314287B2 (en) | ||
| TW201315471A (en) | Compositions and methods for treating cancer using PI3K β inhibitor and mapk pathway inhibitor, including MEK and RAF inhibitors |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| 121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 20830320 Country of ref document: EP Kind code of ref document: A1 |
|
| NENP | Non-entry into the national phase |
Ref country code: DE |
|
| 122 | Ep: pct application non-entry in european phase |
Ref document number: 20830320 Country of ref document: EP Kind code of ref document: A1 |