Immunosuppression modulating compounds
The present invention provides immunosuppression compounds capable of inhibiting the programmed cell death 1 (PD1) signalling pathway. The present invention further provides peptide based compositions for treatment of cancer or treatment of infections via immunopotentiation caused by inhibition of immunosuppressive signaling induced by PD-1, PD-L1, or PD-L2 and therapies using them, immunopotentiative substrates included as the active ingredient. Further, the invention provides an application of the compositions containing the peptide moieties for preventive and/or therapeutic agents for cancer, cancer metastasis, immunodeficiency, an infectious disease or the like and an application of peptide moieties as a testing or diagnostic agent or a research agent for such a disease.
1. A compound of formula (I)
or pharmaceutically acceptable salt thereof;
wherein;
the compound comprises at least 5 amino acids,
A is an amino acid sequence SNTSESF (SEQ ID NO: 4);
B is an amino acid sequence SNTSESF (SEQ ID NO: 4);
Z is
(i) from one to four peptide sequences arranged in any order each being of from three amino acids up to the full length of a mammalian PD1 ectodomain fragment selected from BC loop, D strand, FG loop, G strand, C strand, F strand, C′ strand, C″ strand, C″-D loop, C′ strand to C′—C″ loop, C′ strand to C″ strand or D strand to DE loop;
(ii) G-L-Z′
G is an amino acid sequence of from three amino acids to the full length of a peptide sequence of mammalian PD1 ectodomain fragments from D-strand or is absent;
L is selected from —CO(CH 2 ) n —NH—, or PEG 2-20 KD;
‘n’ is an integer selected from 2 to 10, both inclusive; and
Z′ is one to three peptide sequences arranged in any order each being of from three amino acids up to the full length of a mammalian PD1 ectodomain fragment selected from FG loop and G-strand; or
(iii) from one to four peptide sequences arranged in any order each being of from three amino acids up to the full length of a mammalian PD1 ectodomain fragment selected from D-strand, FG loop and G strand, wherein two or more amino acids of the peptide sequence combine together to form a lactam bond between any of the two fragments or within the fragment;
D is up to two peptide sequences arranged in any order each being of from three amino acids up to the full length of a mammalian PD1 ectodomain fragment selected from BC loop, FG loop, C C′ loop to C′ strand or is absent;
E is up to four peptide sequences arranged in any order each being of from three amino acids up to the full length of a mammalian PD1 ectodomain fragment selected from BC loop, D strand, FG loop, C C′ loop to C′ strand, G strand, FG loop to G strand or is absent;
X is lysine;
X′ is selected from lysine, ornithine, diaminopropionic acid, diaminobutyric acid or olefinic amino acid of formula
which is optionally linked with an additional lysine; or
X′ is absent;
‘m’ is an integer selected from 1 to 6, both inclusive;
R 1 is selected from group consisting of C 2 -C 20 acyl, PEG 2-20 KD moiety; or absent;
R 2 and R 3 are independently selected from group consisting of C 2 -C 20 acyl, PEG 2-20 KD, absent or R a -L′;
R a is selected from biotin or maleimido propionic acid;
L′ is selected from linkers —CO(CH 2 ) n —NH—, —CO(CH 2 —CH 2 —O—) n NH or —COCH 2 (—OCH 2 —CH 2 ) n NH—; and
‘n’ is an integer selected from 2 to 10, both inclusive;
R 4 and R 5 are independently NH 2 , or one or both of R 4 or R 5 are absent with the proviso to the compound of Formula I, that in a compound of Formula I as above defined:
a) up to 5 but not more than 25% of the amino acids may be substituted with other natural or unnatural amino acids;
b) not more than 30% of the amino acids may be omitted;
c) in each said peptide sequence up to 2 amino acids may be added individually at any position;
d) up to 5 but not more than 25% of the peptide bonds may instead be replaced by reduced amide bond (—CH 2 NH—);
e) up to 100% of the amino acids may be D-amino acids;
f) up to 100% of the amino acids may be in reverse order.
2. The compound according to claim 1 , wherein said mammalian PD1 ectodomain fragments are from human, murine, dog, horse or rat PD1.
3. The compound according to claim 1 , wherein Z consists of one said peptide sequence.
4. The compound according to claim 1 , wherein Z is a combination of D strand, FG loop and G strand.
5. The compound according to claim 1 , wherein X is lysine and X′ is absent.
6. The compound according to claim 1 , wherein both X and X′ are lysine.
7. The compound according to claim 1 , wherein R 2 and R 3 are selected from C2-C20 acyl, or R a -L′; wherein,
R a is maleimido propionic acid;
L′ is —COCH 2 (—OCH 2 —CH 2 ) n NH—; and
‘n’ is an integer selected from 2 to 10, both inclusive.
8. The compound according to claim 1 , wherein X′, D, E and R5 are absent and R4 is NH2.
9. The compound according to claim 1 , wherein the compound is selected from:
Comp.
Sequence
No.
Compound
ID
8
SNTSESFK(SNTSESF)FRVTQLAPKAQIKE-NH2
49
13
SNTSESFK (SNTSESF) FRVTQLAPKAQIKE-NH2
54
All D-amino acids
15
Biotin-Ahx-SNTSESFK(SNTSESF)FRVTQLAPKAQIKE-NH2
56
Ahx = 6-aminohexanoic acid
16
C6 lipid-SNTSESFK(SNTSESF)FRVTQLAPKAQIKE-NH2
57
17
Ac-SNTSESFK(SNTSESF)FRVTQLAPKAQIKE-NH2
58
18
SNTSESFK(NH2-[PEG]11-CO-SNTSESF)FRVTQLAPKAQIKE-NH2
59
19
SNTSESFK(SNTSESF)-NH-[PEG]11-CO-)LAPKAQIKE-NH2
60
20
SNTSESFK(CH3(CH2)4COSNTSESF)FRVTQLAPKAQIKE-NH2
61
Lipid in branch
21
CH3(CH2)4CO-SNTSESFK(CH3(CH2)4CO-
62
SNTSESF)FRVTQLAPKAQIKE-NH2
Lipid on both Branch & N-termini
25
SNTSESFK(SNTSESF)FRVTQLAQIKE-NH2
66
29
SNTSESFK(SNTSESF)-NH2
70
30
SNTSESFK(SNTSESF)LAPKAQIKE-NH2
71
31
SNTSESFK(SNTSESF)FRVTQKAQIKE-NH2
72
32
SNTSESFK(SNTSESF)KAQIKE-NH2
73
33
SNTSESFK(SNTSESF)FRVTQLAPKAQIK(CH3(CH2)14CO)E-NH2
74
34
SNTSESFK(SNTSESF)FRVTQLAPK(CH3(CH2)14CO)AQIKE-NH2
75
35
SNTSESFK(SNTSESF)FRVTQK(LAP)KAQIKE-NH2
76
36
SNTSESFK(SNTSESF)FRVTQLAK(PKA)QIKE-NH2
77
40
SNTSESFK(SNTSESF)FK(CH3(CH2)14CO))VTQLAPKAQIKE-NH2
81
Arg in D strand replaced by Lys
41
SNTSESFK(SNTSESF)FRVTQLAP-NH2
82
44
SNTSESFK(SNTSESF)FK(CH3(CH2)6CO)VTQLAPKAQIKE-NH2
85
Arg in D strand replaced by Lys
45
SNTSESFK(SNTSESF)FRVTQLAPK(CH3(CH2)6CO)AQIKE-NH2
86
46
SNTSESFK(SNTSESF)FRVTQLAPKAQIK(CH3(CH2)6CO)E
87
47
SNTSESFK(sntsesf)FRVTQLAPKAQIKE
88
D-amino acids in the branch
48
EKIQAKPALQTVRFK(FSESTNS)FSESTNS-NH2
89
49
EKIQAKPALQTVRFK(FSESTNS)FSESTNS-NH2
90
Retro inverso (All D-amino acid)
50
SNTSESFK(SNTSESF)FRVTQLAPKAQIK(PEG-20KD)E-NH2
91
51
SNTSESFK(SNTSESF)FRVTQLAPKAQIK(PEG-20KD)E-NH2
92
52
SNTSESFK(SNTSESF)FRVTQLAPK(PEG 20 KD)AQIKE-NH2
93
53
SNTSESFK(SNTSESF)FK(mini PEG)VTQLAPKAQIKE-NH2
94
Arg in D strand replaced by Lys
54
SNTSESFK(SNTSESF)FRVTQLAPK(PEG 10KD)AQIKE-NH2
95
55
SNTSESFK(SNTSESF)FRVTQLAPKAQIK(PEG20KD)E-NH2
96
56
SNTSESFK(SNTSESF)FK(CH3CO)VTQLAPKAQIKE-NH2
97
57
SNTSESFK(SNTSESF)FRVTQLAPK(CH3CO)AQIKE-NH2
98
58
SNTSESFK(SNTSESF)FRVTQLAPKAQIK(CH3CO)E-NH2
99
59
SNTSESFK(SNTSESF)FRVTQLAPK(CH3—(CH2)4—CO)AQIKE-NH2
100
60
Biotin-Ahx-SNTSESFK(SNTSESF)FRVTQLAPKAGIKE-NH2
101
All D Amino acids
62
SNTSESFK(SNTSESF)FRVTQLAPE*AQIK*E-NH2
103
Lactam bond formed between asterisked(*) amino acids
63
SNTSESFK(SNTSESF)FK*VTQE*APKAQIKE-NH2
104
Lactam bond formed between asterisked(*) amino acids
64
SNTSESFK(SNTSESF)FRVTE*LAPK*AQIKE
105
Lactam bond formed between asterisked(*) amino acids
65
CH3CO-SNTSESFK(CH3CO-SNTSESF)FRVTQLAPKAQIK(CH3(CH2)6CO)E
106
66
CH3CO-SNTSESFK(CH3CO-SNTSESF)FRVTQLAPKAQIK(CH3(CH2)14CO)E
107
67
CH3CO-SNTSESFK(CH3COSNTSESF)FRVTQLAPKAQIK(CH3(CH2)10CO)E
108
at Lys
68
CH3CO-SNTSESFK(CH3CO-
109
SNTSESF)FRVTQLAPKAQIK(CH3(CH2)4CO)E
77
SNTSESFK(SNTSESF)GIYLCGAISLAPKAQIKE-NH2
118
78
SNTSESFK(SNTSESF)VLNWYRMLAPKAQIKE-NH2
119
83
SNTSESFK(SNTSESF)FRVTQLAPK(MPA-
124
NH—CH2—CH2—O—CH2—CH2—OCO)AQIKE-NH2
84
SNTSESFK(sNTSESF)FRVTQLAPKAQIKE-NH2
125
85
sNTSESFK(SNTSESF)FRVTQLAPKAQIKE-NH2
126
86
sNTSESFK(sNTSESF)FRVTQLAPKAQIKE-NH2
127
87
SNTSESFK(SNTSESF)FRVTQLAPKAQIK(MPA-
128
NH—CH2—CH2—O—CH2—CH2—O—CO)E-NH2
88
SΨ[CH2NH]NTSESFK(SΨ[CH2NH]NTSESF)FRVTQLAPKAQIKE-NH2
129
89
SnTSESFK(SNTSESF)FRVTQLAPKAQIKE-NH2
130
D-Asn at N-terminus
90
SNTSESFK(SnTSESF)FRVTQLAPKAQIKE-NH2
131
D-Asn in the branch
92
SNTSESFK(SNTSESF)FRVTQLAPKAQIKE
133
93
SΨ[CH2NH]NTSESFK(SNTSESF)FRVTQLAPKAQIKE-NH2
134
94
SNTSESFK(SΨ[CH2NH]NTSESF)FRVTQLAPKAQIKE-NH2
135
95
SNΨ[CH2NH]TSESFK(SNTSESF)FRVTQLAPKAQIKE-NH2
136
96
SNTSESFK(SNΨ[CH2NH]TSESF)FRVTQLAPKAQIKE-NH2
137
97
SNΨ[CH2NH]TSESFK(SNΨ[CH2NH]TSESF)FRVTQLAPKAQIKE-NH2
138
98
SNTSESFK(SNTSESF)FRVTQLAPK(CH3(CH2)14CO)AQIKE
139
102
SNTSESFK(SNTSESF)FRVTQLAPK*IAQE*KE-NH2
143
Lactam bond formed between asterisked(*) amino acids
103
SNTSESFK(SNTSESF)K*RVTE*LAPKAQIKE-NH2
144
Lactam bond formed between asterisked(*) amino acids
104
SNTSESFK(SNTSESF)FRK*TQLE*PKAQIKE-NH2
145
Lactam bond formed between asterisked(*) amino acids
105
SNTSESFK(SNTSESF)FRVE*QLAK*PAQIKE-NH2
146
Lactam bond formed between asterisked(*) amino acids.
10. A compound according to claim 1 having formula (Ia)
or pharmaceutically acceptable salt thereof;
wherein;
A is an amino acid sequence SNTSESF (SEQ ID NO: 4);
B is an amino acid sequence SNTSESF (SEQ ID NO: 4);
Z is from one to three peptide sequences arranged in any order each being of from three amino acids up to the full length of a peptide sequence of human or murine PD1 ectodomain fragments selected from D strand, FG loop, G strand, C strand and F strand;
X is lysine;
R 1 is selected from the group consisting of C 2 -C 20 acyl, PEG 2-20 KD moiety; or absent;
R 2 and R 3 are independently selected from group consisting of C 2 -C 20 acyl, PEG 2-20 KD, absent or R a -L′;
R a is selected from biotin or maleimido propionic acid;
L′ is selected from linkers —CO(CH 2 ) n —NH—, —CO(CH 2 —CH 2 —O—) n NH or —COCH 2 (—OCH 2 —CH 2 ) n NH—; and
‘n’ is an integer selected from 2 to 10, both inclusive; and
R 4 is NH 2 .
11. The compound according to claim 10 , wherein Z is a combination of D strand, FG loop and G strand.
12. The compound according to claim 10 , wherein R 1 , R 2 and R 3 are absent.
13. The compound according to claim 10 , wherein R 3 is C 16 -acyl.
14. The compound according to claim 10 , wherein up to three amino acids are D-amino acids.
15. The compound according to claim 10 , wherein all amino acids are D-amino acids.
16. The compound according to claim 10 , comprising up to three reduced amide bonds (—CH 2 NH—).
17. The compound according to claim 1 , wherein Z is three peptide sequences arrange in any order each being of from three amino acids up to the full length of a mammalian PD1 ectodomain fragment selected from BC loop, D strand, FG loop, G strand, C strand, F strand, C′ strand, C″ strand, C″-D loop, C′ strand to C′C″ loop, C′ strand to C′ strand or D strand to DE loop.
18. A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 , and a pharmaceutically acceptable diluent or carrier.
19. A method of modulating an immune response mediated by PD-1 signaling pathway in a subject, comprising administering to the subject therapeutically effective amount of compound according to claim 1 , such that the immune response in the subject is modulated.
20. A method of inhibiting growth of tumour cells and/or metastasis in a subject, comprising administering to the subject a therapeutically effective amount of compound according to claim 1 , capable of inhibiting the programmed cell death 1 (PD1) signaling pathway.
21. The method of claim 20 , wherein the tumour cells are of a cancer selected from the group consisting of melanoma, renal cancer, prostate cancer, breast cancer, colon cancer and lung cancer.
22. The method of claim 20 , wherein the tumour cells are of a cancer selected from the list consisting of bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemias including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumours of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumour angiogenesis, spinal axis tumour, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, and combinations of said cancers.
23. A method of treating an infectious disease in a subject comprising administering to the subject a therapeutically effective amount of compound according to claim 1 , capable of inhibiting the programmed cell death 1 (PD1) signaling pathway such that the subject is treated for the infectious disease.
24. A method of treating bacterial and viral infections in a subject comprising administering to the subject a therapeutically effective amount of compound according to claim 1 , capable of inhibiting the programmed cell death 1 (PD1) signaling pathway such that the subject is treated for the bacterial and viral infections.