PYRROLOBENZODIAZEPINE-ANTIBODY CONJUGATES
The present disclosure relates to the use of ADCs comprising anti-CD25 antibodies for use in treating disorders characterized by the presence of CD25+ve cells.
1 .- 124 . (canceled)
125 . A method of causing cytotoxicity to a neoplastic CD25−ve cell in the vicinity of a CD25+ve cell, the method comprising use of a conjugate of formula L-(D L ) p , where D L is of formula I or II:
wherein:
L is an antibody (Ab) which is an antibody that binds to CD25;
when there is a double bond present between C2′ and C3′, R 12 is selected from the group consisting of:
(ia) C 5-10 aryl group, optionally substituted by one or more substituents selected from the group comprising: halo, nitro, cyano, ether, carboxy, ester, C 1-7 alkyl, C 3-7 heterocyclyl and bis-oxy-C 1-3 alkylene;
(ib) C 1-5 saturated aliphatic alkyl;
(ic) C 3-6 saturated cycloalkyl;
(id)
wherein each of R 21 , R 22 and R 23 are independently selected from H, C 1-3 saturated alkyl, C 2-3 alkenyl, C 2-3 alkynyl and cyclopropyl, where the total number of carbon atoms in the R 12 group is no more than 5;
(ie)
wherein one of R 25a and R 25b is H and the other is selected from: phenyl, which phenyl is optionally substituted by a group selected from halo, methyl, methoxy; pyridyl; and thiophenyl; and
(if)
where R 24 is selected from: H; C 1-3 saturated alkyl; C 2-3 alkenyl; C 2-3 alkynyl; cyclopropyl; phenyl, which phenyl is optionally substituted by a group selected from halo, methyl, methoxy; pyridyl; and thiophenyl;
when there is a single bond present between C2′ and C3′,
R 12 is
where R 26a and R 26b are independently selected from H, F, C 1-4 saturated alkyl, C 2-3 alkenyl, which alkyl and alkenyl groups are optionally substituted by a group selected from C 1-4 alkyl amido and C 1-4 alkyl ester; or, when one of R 26a and R 26b is H, the other is selected from nitrile and a C 1-4 alkyl ester;
R 6 and R 9 are independently selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NRR′, nitro, Me 3 Sn and halo;
where R and R′ are independently selected from optionally substituted C 1-12 alkyl, C 3-20 heterocyclyl and C 5-20 aryl groups;
R 7 is selected from H, R, OH, OR, SH, SR, NH 2 , NHR, NHRR′, nitro, Me 3 Sn and halo;
R″ is a C 3-12 alkylene group, which chain may be interrupted by one or more heteroatoms, e.g. O, S, NR N2 (where R N2 is H or C 1-4 alkyl), and/or aromatic rings, e.g. benzene or pyridine;
Y and Y′ are selected from O, S, or NH;
R 6′ , R 7′ , R 9′ are selected from the same groups as R 6 , R 7 and R 9 respectively;
[Formula I]
R L1′ is a linker for connection to the antibody (Ab);
R 11a is selected from OH, OR A , where R A is C 1-4 alkyl, and SO z M, where z is 2 or 3 and M is a monovalent pharmaceutically acceptable cation;
R 20 and R 21 either together form a double bond between the nitrogen and carbon atoms to which they are bound or;
R 20 is selected from H and R C , where R C is a capping group;
R 21 is selected from OH, OR A and SO z M;
when there is a double bond present between C2 and C3, R 2 is selected from the group consisting of:
(ia) C 5-10 aryl group, optionally substituted by one or more substituents selected from the group comprising: halo, nitro, cyano, ether, carboxy, ester, C 1-7 alkyl, C 3-7 heterocyclyl and bis-oxy-C 1-3 alkylene;
(ib) C 1-5 saturated aliphatic alkyl;
(ic) C 3-6 saturated cycloalkyl;
(id)
wherein each of R 11 , R 12 and R 13 are independently selected from H, C 1-3 saturated alkyl, C 2-3 alkenyl, C 2-3 alkynyl and cyclopropyl, where the total number of carbon atoms in the R 2 group is no more than 5;
(ie)
wherein one of R 15a and R 15b is H and the other is selected from: phenyl, which phenyl is optionally substituted by a group selected from halo, methyl, methoxy; pyridyl; and thiophenyl; and
(if)
where R 14 is selected from: H; C 1-3 saturated alkyl; C 2-3 alkenyl; C 2-3 alkynyl; cyclopropyl; phenyl, which phenyl is optionally substituted by a group selected from halo, methyl, methoxy; pyridyl; and thiophenyl;
when there is a single bond present between C2 and C3,
R 2 is
where R 16a and R 16b are independently selected from H, F, C 1-4 saturated alkyl, C 2-3 alkenyl, which alkyl and alkenyl groups are optionally substituted by a group selected from C 1-4 alkyl amido and C 1-4 alkyl ester; or, when one of R 16a and R 16b is H, the other is selected from nitrile and a C 1-4 alkyl ester;
[Formula II]
R 22 is of formula IIIa, formula IIIb or formula IIIc:
(a)
where A is a C 5-7 aryl group, and either
(i) Q 1 is a single bond, and Q 2 is selected from a single bond and —Z—(CH 2 ) n —, where Z is selected from a single bond, O, S and NH and n is from 1 to 3; or
(ii) Q 1 is —CH═CH—, and Q 2 is a single bond;
(b)
where;
R C1 , R C2 and R C3 are independently selected from H and unsubstituted C 1-2 alkyl;
(c)
where Q is selected from O—R L2′ , S—R L2′ and NR N —R L2′ , and R N is selected from H, methyl and ethyl
X is selected from the group comprising: O—R L2′ , S—R L2′ , CO 2 —R L2′ , CO—R L2′ , NH—C(═O)—R L2′ , NHNH—R L2′ , CONHNH—R L2′ ,
NR N R L2′ , wherein R N is selected from the group comprising H and C 1-4 alkyl;
R L2′ is a linker for connection to the antibody (Ab);
R 10 and R 11 either together form a double bond between the nitrogen and carbon atoms to which they are bound or;
R 10 is H and R 11 is selected from OH, OR A and SO z M;
R 30 and R 31 either together form a double bond between the nitrogen and carbon atoms to which they are bound or;
R 30 is H and R 31 is selected from OH, OR A and SO z M.
126 . The method according to claim 125 , wherein R 7 is a C 1-4 alkyloxy group.
127 . The method according to claim 125 , wherein Y is O and R″ is C 3-7 alkylene.
128 . The method according to claim 125 , wherein R 6 and R 9 are H.
129 . The method according to claim 125 , wherein there is a double bond between C2′ and C3′, and R 12 is:
(a) a C 5-7 aryl group, which may bear one to three substituent groups selected from methoxy, ethoxy, fluoro, chloro, cyano, bis-oxy-methylene, methyl-piperazinyl, morpholino and methyl-thiophenyl; or
(b) methyl, ethyl or propyl; or
(c) cyclopropyl; or
(d) a group of formula:
wherein the total number of carbon atoms in the R 12 group is no more than 4; or
(e) the group:
or
(f) a group of formula:
wherein R 24 is selected from H and methyl.
130 . The method according to claim 125 , wherein there is a single bond between C2′ and C3′, R12 is
and:
(a) R 26a and R 26b are both H; or
(b) R 26a and R 26b are both methyl; or
(c) one of R 26a and R 26b is H, and the other is selected from C 1-4 saturated alkyl, C 2-3 alkenyl, which alkyl and alkenyl groups are optionally substituted.
[Formula I]
131 . The method according to claim 125 , wherein there is a double bond between C2 and C3, and R 2 is:
(a) a C 5-7 aryl group, which may bear one to three substituent groups selected from methoxy, ethoxy, fluoro, chloro, cyano, bis-oxy-methylene, methyl-piperazinyl, morpholino and methyl-thiophenyl; or
(b) methyl, ethyl or propyl; or
(c) cyclopropyl; or
(d) a group of formula:
wherein the total number of carbon atoms in the R 2 group is no more than 4; or
(e) the group:
or
(f) a group of formula:
wherein R 14 is selected from H and methyl.
132 . The method according to claim 125 , wherein there is a single bond between C2 and C3, R 2 is
and:
(a) R 16a and R 16b are both H; or
(b) R 16a and R 16b are both methyl; or
(c) one of R 16a and R 16b is H, and the other is selected from C 1-4 saturated alkyl, C 2-3 alkenyl, which alkyl and alkenyl groups are optionally substituted.
133 . The method according to claim 125 , wherein R 20 is R C , wherein R C is a group:
where the asterisk indicates the point of attachment to the N10 position, G 2 is a terminating group, L 3 is a covalent bond or a cleavable linker L 1 , L 2 is a covalent bond or together with OC(═O) forms a self-immolative linker.
[Formula II]
134 . The method according to claim 125 , wherein:
(a) R 22 is of formula IIIa, A is phenyl, Q 1 is a single bond and Q 2 is a single bond; or
(b) R 22 is of formula IIIb, and R C1 , R C2 and R C3 are all H; and
X is NH—R L2′ .
135 . The method according to claim 125 , wherein R 6′ , R 7′ , R 9′ , and Y′ are the same as R 6 , R 7 , R 9 , and Y.
136 . The method according to claim 125 , wherein L-R L1′ or L-R L2′ is a group:
where the asterisk indicates the point of attachment to the PBD, Ab is the antibody, L 1 is a cleavable linker, A is a connecting group connecting L 1 to the antibody, L 2 is a covalent bond or together with —OC(═O)— forms a self-immolative linker.
137 . The method of claim 136 , wherein L 1 comprises a dipeptide and the group —X 1 —X 2 — in dipeptide, —NH—X 1 —X 2 —CO—, is selected from:
-Phe-Lys-,
-Val-Ala-,
-Val-Lys-,
-Ala-Lys-,
-Val-Cit-.
138 . The method according to claim 136 , wherein C(═O)O and L 2 together form the group:
where the asterisk indicates the point of attachment to the PBD, the wavy line indicates the point of attachment to the linker L 1 , Y is NH, O, C(═O)NH or C(═O)O, and n is 0 to 3.
139 . A method according to claim 125 , wherein
D L is selected from:
140 . The method according to claim 125 , wherein the antibody is:
an intact antibody;
(ii) humanised, deimmunised or resurfaced; or
(iii) a fully human monoclonal IgG1 antibody, preferably IgG1,κ.
141 . The method according to claim 125 , wherein the antibody:
(i) is selected from: basiliximab; daclizumab; HuMax-TAC;
(ii) comprises: a VH domain comprising a VH CDR1 with the amino acid sequence of SEQ ID NO.3, a VH CDR2 with the amino acid sequence of SEQ ID NO.4, and a VH CDR3 with the amino acid sequence of SEQ ID NO. 5;
(iii) comprises a VH domain having the sequence according to SEQ ID NO. 1;
(iv) comprises: a VL domain comprising a VL CDR1 with the amino acid sequence of SEQ ID NO.6, a VL CDR2 with the amino acid sequence of SEQ ID NO.7, and a VL CDR3 with the amino acid sequence of SEQ ID NO.8;
(v) comprises a VL domain having the sequence according to SEQ ID NO. 2.
142 . The method according to claim 125 , wherein the drug loading (p) of drugs (D) to antibody (Ab) is: an integer from 1 to about 8; or is 1, 2, 3, or 4.
143 . The method according to claim 125 , wherein:
both CD25+ve and CD25−ve cells are neoplastic cells;
(ii) the CD25+ve cell is a tumour infiltrating lymphocyte;
(iii) the neoplastic cells are, or are present in, a non-hematological cancer;
(iv) the neoplastic cells are, or are present in, a solid tumor;
(v) the neoplastic cells are malignant; or the neoplastic cells are metastatic.