TECHNICAL FIELD

[0001] The present invention relates to the treatment

and/or prevention of diseases like capillary leak syndrome (CLS) . In particular, the present invention relates to the field of tumor immunotherapy, more specifically to the prevention of the detrimental sequels of such therapy caused by T-cell cytokine release syndrome elicited by various approaches to tumor immunotherapy. Methods for the prevention of vascular leak triggered, for instance, by T-cell related cytokines in these circumstances are disclosed .

BACKGROUND ART

[0002] Tumors have many mechanisms for immune evasion and for modulating immune response through perturbation of the antigen presenting machinery, DNA repair and replication, tumor microenvironment , inflammation pathways, and checkpoint inhibitors/activators. These immune tolerance mechanisms allow tumors to develop invasive growth.

[0003] A number of methods have been developed to break immune tolerance for tumors allowing the immune system and more specific cytotoxic T-cells to recognize, attack and kill tumor cells. These T-cell-engaging immunotherapies are exciting new approaches and include checkpoint inhibitors such as ipilimumab, a CTLA4 specific monoclonal antibody (Lipson et al . , Clin Cancer Res. 2011; 17: 6958- 6962), pembrolizumab (Deeks, Drugs 2016; 76: 375-386) and nivolumab (Johnson et al . , Ther Adv. Med Oncol 2015; 7: 97-106) specific for PD-1, the bispecific antibody blina- tumomab, specific for CD3 and CD19 (Newman et al . , J. Oncol Pharm Pract 2015; 22: 639-645) and chimeric antigen receptor (CAR) modified T cells targeted to CD19 and other tumor cell specific surface antigens

(WO 2015/6033570, WO 2015/164675, WO 2015/120096, Porter et al. New Engl. J. Med. 2011; 365: 725-733, Grupp et al . New. Engl. J. Med. 2013; 368: 1509-1518). These agents have shown unprecedented remission rates for instance in the relapsed, refractory acute lymphatic leukemia (ALL) setting and in other liquid and solid tumors. However, cytokine release syndrome (CRS) , resulting from the high magnitude of immune activation by these therapies, is the most significant and very common treatment-related toxicity. CRS manifests with fever and malaise and can progress to life-threatening capillary leak with hypoxia and hypotension. For instance, in one study 94% of patients with ALL who were treated with CTL019 (engineered T-cells composed of an anti-CDl9 single chain variable fragment (scFv) , aCD3( activation domain, and a 4-1BB costimula- tory domain) developed CRS. Nine out of 51 treated patients required mechanical ventilation indicative of ARDS and 20 patients developed distributive or cardiogenic shock. Both symptoms are typical consequences of capillary leak (Teachey et al . , Cancer Disc. 2016; 6: 664- 679) . These side effects are typically dose limiting and prevent the exploitation of the full therapeutic potential of these interventions. It is therefore highly desirable to protect a subject treated with tumor immunotherapy from the development of such capillary leak syndrome and thus allow the application of high doses of the tumor therapeutic. Hence, it is an object of the present invention to provide methods and means for treating and/or preventing capillary leak syndrome.

SUMMARY OF THE INVENTION

[0004] The present invention relates to a method for inhibiting, treating and/or preventing a capillary leak syndrome (CLS) or symptoms thereof in a subject treated with a T-cell or NK cell activating agent, said method comprising administering fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof or a physiologically acceptable salt thereof to said subject.

[0005] It turned surprisingly out that the administration of fibrin-derived peptide B Beta 15-42 and/or derivatives thereof can be used to optimize the dosing of T- cell and NK cell activating agents and to exploit their anti-tumor effect to its full extent.

[0006] Another aspect of the present invention relates to fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof for use in inhibiting, treating and/or preventing a capillary leak syndrome in a subject treated with a T-cell or NK cell activating agent.

[0007] A further aspect of the present invention relates to a composition comprising fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof as defined herein for use in inhibiting, treating and/or preventing a capillary leak syndrome in a subject treated with a T- cell or NK cell activating agent

DESCRIPTION OF EMBODIMENTS

[0008] The disruption of the adherence junction between endothelial cells is the hallmark of capillary leak syndrome. This disruption is caused by a rearrangement of the actin skeleton of the endothelial cell and the formation of stress fibres. This disruption can be caused by many effector molecules including cytokines released after activation of T-cells. Symptoms of capillary leak syndrome include elevated hematocrit or hemoglobin readings (with hematocrit levels >40% due to the leak of plasma) , very low blood pressure (profound arterial hypotension, with systolic blood pressure levels <90 mm Hg) , albumin deficiency (hypoalbuminemia measuring <3.0 g/dL) , partial or generalized edema, cold extremities and a paraprotein in the blood (an MGUS in approximately 80% of cases) .

[0009] Herein, it could be surprisingly shown that the

treatment with fibrin-derived peptide B Beta 15-42 and/or peptides, peptide derivatives or peptidomimetics , preferably of the general formulae (I-VI) , act synergistically with the treatment of various cancers with immunologically active agents by allowing to maximize the dose of these agents for optimal anti-tumour efficacy, while avoiding the potentially lethal capillary leak syndrome, preferably caused by stimulation of T-cell related cytokines .

[0010] Therefore, the present invention relates to a method for inhibiting, treating and/or preventing a capillary leak syndrome or symptoms thereof in a subject treated with a T-cell or NK cell activating agent, said method comprising administering fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof or a physiologically acceptable salt thereof to said subject. Fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof is administered to said subject in a therapeutically effective amount.

[0011] The present invention relates in another aspect also to a fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof for use in inhibiting, treating and/or preventing a capillary leak syndrome in a subject treated with a T-cell or NK cell activating agent.

[0012] The instant invention encompasses methods for inhibiting, treating and/or preventing a cytokine related vascular leak syndrome precipitated by treatment of a cancer patient with an agent that stimulates T-cells to attack and kill tumor cells.

[0013] "Inhibiting" a capillary leak syndrome, as used

herein, means that the administration of fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof stops or reduces significantly the progress and symptoms of the disorder. The inhibition/reduction may be at least 10%, preferably at least 20%, more preferably at least 50%, compared to a subject suffering from capillary leak syndrome who does not receive therapeutically amounts of fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof. If the capillary leak syndrome is a result of an increased cytokine secretion within the subject's body (e.g. due to the administration of T-cell activating agents like antibodies) , "inhibition" may also mean that the effect of an increased cytokine secretion is inhibited and/or reduced. This inhibi- tion can for instance be measured by a reduction in ex- travascular lung water using the Picco instrument, by a reduction of the required dose of catecholamines to alleviated critical hypotension or parameter measuring renal function, e.g. creatinin clearance, as well as reduction of cytokine release.

[0014] As used herein, the terms "treating" and "treatment" refer to anything that promotes or enhances the well-being of the subject with respect to the medical treatment of his or her condition. This includes, but is not limited to, a reduction in the frequency or severity of the signs or symptoms of capillary leak syndrome. Since in a preferred embodiment of the present invention fibrin-derived peptide B Beta 15-42 and/or derivatives thereof can be used in subjects suffering from capillary leak syndrome caused by T-cell activating agents like antibodies making the disorder in most or even all cases reversible "treatment" may refer to a reduction of the disorder and/or symptoms thereof to an extent of at least 10%, more preferably at least 20%, preferably at least 30%.

[0015] The terms "prevention" and "preventing", as used

herein, refer to the administration or application of fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof to a subject for the purpose of blocking the onset of capillary leak syndrome. For example, a subject that does not yet meet the clinical criteria or exhibit the symptoms of capillary leak syndrome, but has been treated with a T-cell or NK cell activating agent may be administered B Beta 15-42 and/or at least one derivative thereof to prevent or delay the onset of capillary leak syndrome or reduce the severity of the condition .

[0016] The term "subject", as used herein, refers to mammals and most preferably to humans.

[0017] To a "subject treated with a T-cell or NK cell activating agent" at least one T-cell or NK cell activating agent is administered in an amount effective to "activate" T-cells and NK cells. [0018] The term "T-cell activating agent", as used herein, is meant to indicate any agent capable of inducing a T- cell or a modified T-cell to enter from a resting state into a state in which the T-cell undergoes the cell cycle or at least parts of it, recognizes a target tumor cell, releases cytokines and is able to kill the target cell. Such agents induce in T cells or modified T cells those processes required for an immune response.

[0019] The term "NK cell activating agent", as used herein, is meant to indicate any agent capable of increasing the activity of NK cells. NK cells are usually active cells which show cytotoxic activity and are responsible among others for clearance of cells infected by viruses, senescent cells and malignantly transformed cells (i.e. cancer cells) . The capability of agents to activate NK cells according to the present invention can be tested using methods known in the art (e.g. Nederby L et al . J Immunol Meth 458 (2018) : 21-25) .

[0020] A "derivative" of fibrin-derived peptide B Beta 15-42 may comprise one or more amino acid changes compared to the wild-type peptide. Preferably the first four amino acid residues of the derivative are identical to the wild-type peptide. The N-terminal end of the derivative is preferably a free amino group whereas the C-terminal end may be modified by the addition of organic groups like polyethylene glycol (PEG) . The derivative of the present invention may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acid exchanges compared to the wild-type peptide. The derivative may be a monomer, dimer, trimer or tetramer of a modified or wild-type fibrin-derived peptide B Beta 15-42 whereby the monomers of the aforementioned multimers are linked to each other by disulphide bonds of cysteine residues, for instance, or other chemical moieties forming a bridge between the monomers. The derivatives of the present invention exhibit properties for inhibiting, treating and/or preventing a capillary leak syndrome in a subject treated with a T-cell or NK cell activating agent. [0021] Fibrin-derived peptide B Beta 15-42 and at least one derivative thereof can be administered in the form of physiologically acceptable nontoxic salts, such as acid addition salts or metal complexes, e.g., with zinc, iron or the like. Illustrative of such acid addition salts are hydrochloride, hydrobromide, sulphate, phosphate, male- ate, acetate, citrate, benzoate, succinate, malate, ascorbate, tartrate and the like.

[0022] According to a preferred embodiment of the present invention the T-cell or NK cell activating agent is administered to a subject in need of an immunotherapy, in particular of a tumor therapy.

[0023] In tumor therapy T-cell or NK cell activating agents are often used. Such agents may include chemotherapeutics as well as antibodies resulting in a T-cell or NK cell activation. These agents are known to result in a significant increase in cytokine secretion, for instance, which may lead to a cytokine "storm". The increased cytokine secretion may result in capillary leak syndrome.

[0024] According to another preferred embodiment of the present invention the T-cell or NK cell activating agent is administered simultaneously or successively to said fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof.

[0025] The fibrin-derived peptide B Beta 15-42 and/or at

least one derivative thereof of the current invention can be co-administered (sequentially or simultaneously) with a tumor immunotherapy agent and/or at least one other agent to prevent and/or treat the CLS symptoms. Tumor immunotherapy agents may include but not only IL 6-blocking antibodies (e.g. tocilizumab) or a corticosteroid.

[0026] "Simultaneous" administration of active ingredients (e.g. a T-cell and/or NK cell activating and fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof) , as used herein, means that at least two active ingredients are administered at the same time, preferably together, to a subject. [0027] Alternatively, the active ingredients of the present invention (i.e. a T-cell and/or NK cell activating and fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof) may be administered to a subject "successively" or "sequentially". This means that one active ingredient is administered to a subject in a first point in time and another active ingredient in a second point in time. The time in between administration of the active ingredients may vary from 1 min to 72 hrs, preferably from 1 min to 48 hrs, more preferably from 5 min to 24 hrs.

[0028] According to a preferred embodiment of the present invention the fibrin-derived peptide B Beta 15-42 consists of amino acid sequence GHRPLDKKREEAPSLRPAPPPISGGGYR (SEQ ID No. 1) .

[0029] It is particularly preferred to use a wild-type fibrin-derived peptide B Beta 15-42 in the method of the present invention.

[0030] Derivatives of fibrin-derived peptide B Beta 15-42 are known in the art. For instance, WO 2007/095659, WO 2007/095660, WO 2007/095661, WO 2009/137850,

WO 2009/137851 and WO 2009/137852 disclose derivatives of fibrin-derived peptide B Beta 15-42, which can be used in the method of the present invention.

[0031] According to a further preferred embodiment of the present invention the fibrin-derived peptide B Beta 15-42 derivative has a general formula selected from the group consisting of

H2N-GHRPX1X2X3X4X5X6X7X8PX9X10X11PX12PPPX13X14X15X16GYR-X 17 ( I ) , (SEQ ID No. 2)

H2N-GHRPX1X2X3X4X5X6X7X8PX9X10X11PX12PPPX13X14X15CGYR Xi8

S

I (Ha) S

H2N-GHRPX1X2X3X4X5X6X7X8PX9X10X11PX12PPPX13X14X15CGYR Xi8,

(SEQ ID No. 3) H2N-GHRPX1X2X3X4X5X6X7X8PX9X10X11PX12PPPX13X14C X15GYR Xi8

S

I (lib) s

H2N-GHRPX1X2X3X4X5X6X7X8PX9X10X11PX12PPPX13X14CX15GYR X18,

(SEQ ID No. 4)

H2N-GHRPX1X2X3X4X5X6X7X8PX9X10X11PX12PPPX13X14X15X16GYR-K - Xl9

B (III)

H2N-GHRPX1X2X3X4X5X6X7X8PX9X10X11PX12PPPX13X14X15X16GYR-K - Xl9, (SEQ ID No. 5)

H2N-GHRPX1X8PX9X10X11PX12PPPX13X14X15X16B (1) B (2) B (3) -Xi7

(IV) ,

(SEQ ID No. 6)

H2N-GHRPX1X2X3X4X5X6X7X8PX9X10X11PX12PPPX13X14X15X16B (1) B (2) B (3) -Xi7 (V)

(SEQ ID No. 7) and

H ₂ N-GHRPX ₂ 0X2lX22-p-X23X24X25X26X27X28X29-Xl7 (VI) ,

(SEQ ID No. 8) wherein

Xl, X2, X3, X4, X5, Χδ, X7, Χδ, Χθ, XlO, Xll, Xl2, X13, Xl4, Xl5 and Xi6 are independently selected from the group of amino acid residues,

Xi7 is

a) ORi, wherein Ri is hydrogen or a C1-C10 alkyl group, or b) NR2R3 , wherein R2 and R3 are independently hydrogen, a Ci-ClO alkyl group, or c) a residue -PEG5-6OK, wherein the PEG-residue is linked to the N atom via a spacer, or d) a residue NH-Y1-Z-PEG5-6OK, wherein Yi is a chemical bond or an amino acid residue selected from the group consisting of S, C, K and R, and Z is a spacer by way of which a polyethylene glycol (PEG) - residue is linked, or

a) ORi, wherein Ri is hydrogen or a C1-C10 alkyl group, or b) NR2R3 , wherein R2 and R3 are independently hydrogen, a C1-C10 alkyl group, or c) a residue -PEG5-6OK, wherein the PEG-residue is linked to the N atom via a spacer, or d) a residue NH-Y1-Z-PEG5-6OK, wherein Yi is a chemical bond or an amino acid residue selected from the group consisting of S, C, K and R, and Z is a spacer by way of which a polyethylene glycol (PEG) - residue is linked, or e) a residue -PEG5-6OK-CO-NR4R5, wherein R4 and R5 are independently hydrogen or a C1-C10 alkyl group, or f) a residue NH-CH (CONH2 ) - (CH ₂ ) 4-NH-CO-Y2-PEG5-6OK, wherein Y2 is an oxygen atom or an NH group, H or NH ₂ , X20, X21, X22, X23, X24, X25 and X26 are independently se ^¬ lected from the group consisting amino acid residues,

X27, X28 and X29 are independently selected from the group consisting amino acid residues or are independently a single bond

B is -CO- (CH ₂ ) m-Y3- (CH ₂ ) m-CO- bound via the CO group to the ε amino group of amino acid residue K, wherein m is an integer from 1 to 4 and Y3 is

-N-CO- (CH ₂ ) n-NH-CO-Z-PEG ₅ -60K or

-N-CO- (CH ₂ ) n-NH-CO-CH- (CH ₂ ) 4-NH-CO-Z-PEG5-6OK

NH-CO-Z-PEG5-6OK, wherein n is an integer from 1 to 4 and Z is NH or 0,

B(l) is a chemical bond or G,

B(2) is a chemical bond or Y,

B(3) is a chemical bond or R,

β is an amino acid residue or a peptidomimetic element, wherein said amino acid residue is selected from the group consisting of L-proline, D-proline, L-hy- droxyproline, D-hydroxyproline, L- (0-benzyl) -hy- droxyproline, D- (O-benzyl) -hydroxyproline, L- (0- tert . butyl) -hydroxyproline, 4- (0-2-naphtyl) -hydroxyproline, 4- (0-2-naphtyl-methyl) - hydroxyproline, 4- (0-phenyl) -hydroxyproline, 4- ( 4-phenyl-ben- zyl) -proline, cis-3- phenyl-proline, cis-4-phenyl- proline, trans-4-phenyl-proline, cis-5-phenyl-pro- line, trans-5-phenyl-proline, 4-benzyl-proline, 4- bromobenzyl-proline, 4-cyclohexyl- proline, 4- fluor-proline, L-tetrahydroisoquinoline-2-carbox- ylic acid (L-Tic) , all diastereomers of octahydro- indole-2-carboxylic acid (Oic) , and all diastere- omers of 1-aza-bicyclo [ 3 , 3 , 0 ] octane-2-carboxylic acid, and wherein said peptidomimetic element is selected from the group consisting of cis-2-amino- cyclopentane carboxylic acid (cis-Acpc) , (1R, 2R) -

(2-aminocyclopentane carboxylic acid ( (1R, 2R) - Acpc), (IS, 2S) - (2-aminocyclopentane carboxylic acid

( (IS, 2S) -Acpc) , 1-aminomethyl-cyclohexane acetic acid (1-Achc) , 3-amino-l-carboxymethyl-pyridin-2- one (Acpo) , 1-amino-cyclobutane-carboxylic acid (1- Acbc) , 1-amino-cyclohexane-carboxylic acid (1- Achc) , cis-4-amino-cyclohexane-acetic acid (4- Acha) , (1R, 2R) -2-aminocyclohexane carboxylic acid

( ( 1R, 2R) -Ache) , (1R, 2S) -2-aminocyclohexane carboxylic acid ( (1R, 2S) -Ache) , (IS, 2R) -2-aminocyclohexane carboxylic acid ((IS, 2R)-Achc), (IS, 2S) -2-aminocyclohexane carboxylic acid ( (IS, 2S) -Ache) , 1- amino-cyclopentane carboxylic acid (1-Acpec) , 1- amino-cyclopropane carboxylic acid (1-Acprc) , 4- (2- aminoethyl) -6-dibenzofuranpropionic acid (Aedfp) ,

(R, S) -1-aminoindane-l-carboxylic acid (1-Aic) , 2- aminoindane-2-carboxylic acid (2-Aic) , 2'-(aminome- thyl) -biphenyl-2-carboxylic acid (Ambc) , 2-aminome- thyl-phenylacetic acid (Ampa) , 3-amino-2-naphthoic acid (Anc) , 4-amino-tetrahydropyran-4-carboxylic acid (Atpc) , (R, S) -2-aminotetraline-2-carboxylic acid (2-Atc) , (2S, 6S, 9S) -6-amino-2-carboxymethyl- 3, 8-diazabicyclo- [4,3,0] -nonane-1, 4-dione (Acdn) ,

(R) -3-amino-5-carboxymethyl-2 , 3-dihydro-l, 5-benzo- thiazepin-4 ( 5H) -one (Acbt) , (S) -3-amino-5-carbox- ymethyl-2, 3-dihydro-l, 5-benzoxazepin-4 (5H) -one

(Acbo) , (R, S) -3-amino-l-carboxymethyl-2 , 3, 4, 5-tet- rahydro-lH- [1] -benzazepin-2-one (1-Acmb) , (S)-4- amino-2-carboxymethyl-l , 3,4, 5-tetrahydro-2H- [2 ] - benzazepin-3-one (2-Acmb) , (R, S) -3-amino-l-carbox- ymethyl-valerolactame (Acmv) , 3- (2-aminoethyl) -1- carboxymethyl-quinazoline-2 , 4-dione (Acq), (2S,5S)- 5-amino-l, 2,4,5,6, 7-hexahydro-azepino [3, 2, 1-hi] - indole-4-one-2-carboxylic acid (Haic) , ( R, S)-3- amino-N-l-carboxymethyl-2-oxo-5-cyclohexyl- 1, 4- benzodiazepine (Accb) , ( R, S) -3-amino-N-l-carbox- ymethyl-2-oxo-5-phenyl-l , 4-benzodiazepine (Acpb) , (2S, llaS) -2-amino-lO-carboxymethyl-l , 2, 3, lla-tetra- hydro-lOH-pyrrolo [2 , 1-c] [1,4] -benzodiazepine-5 , 11- dione (PBD) , (2S, 3 ' S) -2- (4 ' - (3 ' -benzyl-2 ' -oxo-pi- perazin-l-yl) ) -3-phenyl- propionic acid (Bppp) , 3- carboxymethyl-l-phenyl- 1,3, 8-tria- zaspiro [ 4.5 ] decan-4-one (Cptd) , ( R, S)-3 -amino-9- Boc-1, 2, 3, 4-tetrahydro-carbazole-3-carboxylic acid (The) , 3-exo-amino-bicyclo [2.2.1] heptane-2-exo-car- boxylic acid (Abhc) , (3S) -3-Amino-l-carboxymethyl- caprolactam (Accl) , (S, S) - (ProLeu) spirolactamePhe (PLSP) and 2-Oxo-3-amino-7-thia-l-azabicy- clo [ 4.3.0 ] nonane- 9-carboxylic acid (BTD) .

[0032] According to a preferred embodiment of the present invention

Xi5 or Xi6 of formula (I) is an amino acid selected from the group consisting of C and K, which is linked to residue Z-PEG5-6OK via the heteroatom in the side chain, and wherein

Xi7 of formula (I) is

a) ORi , wherein Ri is hydrogen or a Ci-Cio-alkyl group, or b) NR2R3, wherein R2 and R3 are independently hydrogen or a C1-C10 alkyl group.

[0033] According to another preferred embodiment of the present invention

Xi , Xg , X10, Xi4, X20 and X23 are independently L, I, S, M or A,

X2, Χβ, X7 and X21 are independently E or D,

X3, X4, X5 X11 and X22 are independently R or K

Xs X12, X24, X25 and X26 are independently A, G, S, or

L, Xi3 is I, L or V,

Xi5 and Xi6 of formula (Ila) , (lib) are independently G, A, S, or C,

X27 is G, A or L,

X28 is Y, F, H or a single chemical bond and X29 is R, K or a single chemical bond.

[0034] According to a preferred embodiment of the present invention a compound of formula I is preferably selected from the group consisting of GHRPLDKKREEAPSLRPAPPPISGG- GYR-NH2 (SEQ ID No. 9), GHRPLDKKREEAPSLRPAPPPISGGGYRC- (S- CH2-CO-NH-PEG20K ) -OH (SEQ ID No. 10), GHRPLDKKREEAPSLRPAPPPISGGGYRC- (S-CH2-CO-NH-PEG20K ) -amid (SEQ ID No. 11) and GHRPLDKKREEAPSLRPAPPPISGC- ( S-CH ₂ -CO-NH-PEG ₂ OK) -GYR-amid (SEQ ID No. 12) .

[0035] According to another preferred embodiment of the present invention a compound of formula Ila is preferably (GHRPLDKKREEAPSLRPAPPPISGCGYR) 2 a Cys25-Cys25 homodimeric cysteine peptide (SEQ ID No. 13).

[0036] According to another preferred embodiment of the present invention a compound of formula lib is

(GHRPLDKKREEAPSLRPAPPPISCGGYR) 2 a Cys24-Cys24 homodimeric cysteine peptide (SEQ ID No. 14) .

[0037] According to a further preferred embodiment of the present invention a compound of the formula III is:

H2N-GHRPLDKKREEAPSLRPAPPPISGCGYRX17 CH2-CO-NH- PEG5-30K (P a)

H ₂ N- GHRPLDKKREEAPS LRP APPPISGCG YRX 17

(SEQ ID No. 15) or H2N-GHRPLDKKREEAPSLRPAPPPISCGGYRX17

S-CH

/ CH- CH2-CO-NH- PEG5-30K (IVb) S- CH ₂ ^X

H ₂ N- GHRPLDKKREEAPS LRP APPPISCGG YRX 17

(SEQ ID No. 16)

[0038] In another preferred embodiment of the present invention a compound of formula IV is selected from the group consisting of GHRPLAPSLRPAPPPISGGGYR -OH (SEQ ID No. 17), GHRPLAPSLRPAPPPISGGGYR -NH ₂ (SEQ ID No. 18),

GHRPLAPSLRPAPPPISGGGYRC- (S-succinimide-PEG ₂ 0K ) -OH (SEQ ID No. 19) and GHRPLAPSLRPAPPPISGGGYRC- (S-succinimido- PEG20K ) -amide (SEQ ID No. 20).

[0039] In another preferred embodiment of the present invention a compound of formula V is selected from the group consisting of GHRPLDKKREEAPSLRPAPPPISGG-OH (SEQ ID No. 21), GHRPLDKKREEAPSLRPAPPPISGG-NH2 (SEQ ID No. 22),

GHRPLDKKREEAPSLRPAPPPISGGG-OH (SEQ ID No. 23) and

GHRPLDKKREEAPSLRPAPPPISGGG-NH2 (SEQ ID No. 24) .

[0040] In a further preferred embodiment of the present invention a compound of formula VI is selected from the group consisting of GHRPLDK- (IS, 2R) Achc-ISGGGYR (SEQ ID No. 25), GHRPLDK-Acdn-ISGGGYR (SEQ ID No. 26), GHRPLDK- (cis-4-Acha) -ISGGGYR (SEQ ID No. 27) and GHRPLDK-Haic-IS- GGGYR (SEQ ID No . 28) .

[0041] According to a preferred embodiment of the present invention it is possible to administer fibrin-derived peptide B Beta 15-42 as the sole active ingredient or in combination with at least one derivative thereof. Of course, it is also possible to administer at least one derivative of fibrin-derived peptide B Beta 15-42 as the sole active ingredient or in combination with fibrin-derived peptide B Beta 15-42.

[0042] Fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof may be administered to a subject in need thereof by any means known to be suitable to those skilled in the art, including intravenous, subcutaneous, intramuscular and mucosal administration. A formulation may thus be formulated for administration via the intravenous, subcutaneous, intramuscular and mucosal route. The mucosal route may be exemplified by, but is not limited to, the pulmonary, nasal, sublingual or buccal route.

[0043] For intravenous, subcutaneous or intramuscular administration, the formulation may be provided as a sterile solution, suspension or an emulsion. The formulation may be applied by means of an injection or an infusion. For mucosal administration, the formulation may be provided as an aqueous spray and may be applied directly by means of a spray container or an inhalator. Alternatively the formulation may be administered to the mucosa as an aqueous gel.

[0044] Fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof may be provided in a liquid or solid form. Both components may be part of a lyophilisate (solid form) which can be combined with a buffer or a physiological salt solution prior administration to a subject. Alternatively, the components may be part of a liquid formulation which may include oils, polymers, vitamins, carbohydrates, amino acids, salts, buffers, albumin, surfactants, or bulking agents. Exemplary carbohydrates include sugar or sugar alcohols such as mono-, di- or polysaccharides, or water-soluble glucans . The saccharides or glucans can include fructose, dextrose, lactose, glucose, mannose, sorbose, xylose, maltose, sucrose, dex- tran, pullulan, dextrin, alpha- and beta-cyclodextrin, soluble starch, hydroxethyl starch and carboxymethyl- cellulose, or mixtures thereof. "Sugar alcohol" is defined as a C4 to Ce hydrocarbon having an -OH group and includes galactitol, inositol, mannitol, xylitol, sorbitol, glycerol, and arabitol. These sugars or sugar alcohols mentioned above may be used individually or in combination. In some embodiments, one can use a buffer in the composition to minimize pH changes in the solution before lyophilization or after reconstitution . Any physiological buffer may be used, but in some cases can be selected form citrate, phosphate, succinate, and glutamate buffers or mixtures thereof.

[0045] The inventive compounds show an effect at a dose

ranging from 0.001 mg/kg body weight to 500 mg/kg body weight, preferably at a dose ranging from 0.1 mg/kg to 50 mg/kg .

[0046] The administration of fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof allows to increase the dose of T-cell and/or NK cell activating agents 1.2-fold up to 10-fold, preferably, 1.5 up to 5 fold, more preferably 1.5 up to 3 fold, because fibrin- derived peptide B Beta 15-42 and its derivatives reduce the side effects caused by both activating agents. Hence, in a preferred embodiment of the present invention the dose of said T-cell or NK cell activating agent administered to a subject within said immunotherapy can be increased when administered simultaneously or successively with fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof compared to the administration of the T-cell or NK cell activating agent without fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof .

[0047] According to a preferred embodiment of the present invention said T-cell activating agent is a checkpoint inhibitor .

[0048] Checkpoint inhibitor therapy is a form of cancer

treatment immunotherapy. The therapy uses immune checkpoints which affect immune system functioning. Immune checkpoints can be stimulatory or inhibitory. Tumors can use these checkpoints to protect themselves from immune system attacks . Checkpoint therapy can block inhibitory checkpoints, restoring immune system function.

[0049] According to a preferred embodiment of the present invention said checkpoint inhibitor is selected from the group consisting of a CTLA4 antagonist, a PDl antagonist and PDL-1 antagonist. [0050] In one embodiment of the invention, fibrin-derived peptide B Beta 15-42 and/or peptides, peptide derivatives or peptidomimetics of the general formulae (I-VI) are administered simultaneously and/or sequentially with a checkpoint inhibitor, exemplified but not limited to an inhibitor to CTLA4 (e.g. ipilimumab) or PD-1 or PDL-1 (e.g. pembrolizumab or nivolumab) .

[0051] In another embodiment of the present inventionthe T- cell activating agent is a bi-specific antibody engaging T-cells to tumor cells comprising at least one binding site for activating molecules on the T-cell surface and at least one binding site for a tumor specific antigen on the tumor cell.

[0052] According to further preferred embodiment of the present invention the T-cell activating agent is a bi-specific antibody with binding sites for CD3 and CD19.

[0053] In another embodiment of the invention, fibrin-derived peptde B Beta 15-42 and/or peptides, peptide derivatives or peptidomimetics of the general formulae (I-VI) are administered simultaneously and/or sequentially with an agent capable of engaging T-cells with tumor cells, exemplified but not limited to a bi-specific antibody targeting CD3 and CD19 (e.g. blinatumomab) .

[0054] According to another embodiment of the present invention the tumor specific antigen is selected from the group consisting of CD19, CD22, BCMA, CD123, Her2, EG- FRvIII, PSCA and GD2.

[0055] According to a further embodiment of the present invention the activating molecule in the T-cell is selected from the group consisting of CD28, 4-1BB, OX40 and ICOS.

[0056] In another embodiment of the invention, fibrin-derived peptide B Beta 15-42 and/or peptides, peptide derivatives or peptidomimetics of the general formulae (I- VI) are administered simultaneously and/or sequentially with engineered T-cells carrying a chimeric antigen receptor, which binds to tumor cells, and co-activation receptors required for activation of the T-cell. This is exemplified by but not limited to CTL019, engineered T- cells composed of an anti-CDl9 single chain variable fragment (scFv) , aCD3( activation domain, and a 4-1BB costimulatory domain.

[0057] In a particular embodiment, a dose of the tumor immunotherapy agent is given in conjunction with fibrin-derived peptide B Beta 15-42 and/or peptides, peptide derivatives or peptidomimetics of the general formulae (I- VI) , which is higher than the dose that can be safely applied without this adjunct treatment because of the elic- itation of potentially lethal capillary leak syndrome.

[0058] Fibrin-derived peptide BBeta 15-42 and/or peptides, peptide derivatives or peptidomimetics of the general formulae (I-VI) can be given in association with a suitable pharmaceutical carrier and can be dosed by various application route, e.g. by bolus intravenous or subcutaneous injection or by continuous intravenous infusion.

[0059] Dosage of fibrin derived peptide B Beta 15-42 or peptides, peptide derivatives or peptidomimetics of the general formulae (I-VI) may vary with the particular need of the patient treated. Dosage units may be increased or decreased based on the weight of the patient. The pharmaceutical preparation comprising the agent may be administered at appropriate intervals until the pathological symptoms are reduced or alleviated. These intervals in a particular case will normally depend on the condition of the patient.

[0060] While certain of the preferred embodiments of the present invention have been described above, it is not intended that the invention is limited to such embodiments. Various modifications may be made thereto without departing from the scope and spirit of the present invention , as set forth in the following claims.

[0061] Another aspect of the present invention relates to fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof for use in inhibiting, treating and/or preventing a capillary leak syndrome in a subject treated with a T-cell or NK cell activating agent. [0062] Yet another aspect of the present invention relates to a composition comprising fibrin-derived peptide B Beta 15-42 and/or at least one derivative thereof as defined herein for use in inhibiting, treating and/or preventing a capillary leak syndrome in a subject treated with a T- cell or NK cell activating agent.

[0063] The composition of the present invention may comprise pharmaceutically acceptable excipients which are usually used to manufacture compositions suited to be administered in a form as defined herein.

[0064] The present invention is further illustrated by the following example without being restricted thereto.

EXAMPLES

[0065] Vascular permeability assay

[0066] Electrical impedance of endothelial cells is measured using the ECIS system (Applied Biophysics) . Human umbilical cord endothelial cells are seeded on gelatin-coated plates at a density of 12000 cells per well and grown to confluence at 37°C and 5% CO2. After resistance at 4000Hz reaches a stable plateau >1000Ω, CD3 positive T cells isolated from peripheral blood and pre-stimulated with phorbol 12-myristate 13-acetate (PMA) (10 ng/ml) and Ionomycin ( 0.4 μΜ) for 6h are added at a concentration of 12000 per well together with FX06 (GHRPLDKKREEAPSLRPAP- PPISGGGYR; SEQ ID No. 1) at a concentrations of 10 and 60 g/ml or random peptide (DRGAPAHRPPRGPISGRSTPEKEKLLPG; SEQ ID No. 29) . Permeability is determined by continuously monitoring electrical impedance at 250 Hz for 48h at 37°C and 5% C0 ₂ (Arterioscler Thromb Vase Biol.2016 Apr; 36 (4) : 647-54. doi : 10.1161/ATVBAHA.115.307032 ) . Results are shown as an increase in resistance (=reduced permeability) in percent compared to the effect of random peptide. n=10, p<0.05 for both concentrations of FX06 compared to random peptide (ANOVA) .

% increase in resistance compared to random peptide time after start of the

coculture 6h 12h 24h

FX06 10 g 23+/-12 33+/-21 34+/-12

FX06 60 g 49+/-20 70+/-17 85+/-32