WO2021185842A1

WO2021185842A1 - Use of substance and pharmaceutical composition thereof, and medical treatments or uses thereof

Info

Publication number: WO2021185842A1
Application number: PCT/EP2021/056694
Authority: WO
Inventors: Tim MELANDER BOWDEN; Thomas Engstrand
Original assignee: Pvac Medical Technologies Ltd
Priority date: 2020-03-16
Filing date: 2021-03-16
Publication date: 2021-09-23
Also published as: SE544131C2; SE2050286A1

Abstract

A substance comprising a carrier exhibiting at least one scavenger structure, said scavenger structure comprising a nucleophilic centre complying with the formula X¹(-R''-)(-R')_mH_n (formula I) wherein a) X¹ is a single-bonded heteroatom selected amongst N, O and S and exhibits a free electron pair, b) m is 0 or 1 and n is 1 or 2 with the sum of m plus n being 2 for X¹ = N and 1 for X¹ = S and O, c) -R''- is a bivalent organic group providing attachment to the carrier via one of its free valences and direct attachment to the heteroatom X¹ at the other one of its free valences, and d) R'- is a monovalent organic group directly attached to the heteroatom X¹ via its free valence for use as a medicament for blocking or inhibiting semicarbazide sensitive amine oxidase (SSAO), preferably blocking or inhibiting SSAO expressed and produced by vascular smooth muscle cells and/or endothelial cells and/or soluble SSAO.

Description

USE OF SUBSTANCE AND PHARMACEUTICAL COMPOSITION THEREOF, AND MEDICAL TREATMENTS OR USES THEREOF

TECHNICAL FIELD The present invention relates to the use of a pharmaceutical composition comprising a substance for prevention and treatment, as well as a method of prevention and treatment pulmonary inflammation diabetes-related complications comprising administering said pharmaceutical composition. BACKGROUND TECHNOLOGY

Semicarbize amine oxidase (SSAO or VAP-1) is an enzyme expressed for example by endothelial cells and smooth muscle cells in blood vessels. SSAO is normally found in intracellular granulae. The enzyme translocates to the cell membrane upon activation and can be solubilized by proteolytic cleavage from the membrane. SSAO exerts dual functions including leukocyte recruitment and also functions as ectoenzyme by the oxidation and conversion of primary amines to aldehydes, hydrogen peroxide and ammonium. There are SSAO inhibitors but some have shown to be toxic or are not very efficient.

In WO 2009108100, inter alia carbazate-functionalized polyvinyl alcohol (PVAC) and related substances was used to treat inflammatory conditions caused by various antigens. The inventors later discovered that the same substances had an antibacterial effect in that they affected the adhesion of the bacteria to surfaces and/or the growth of the bacteria. Thus, the substances may thus also be used to prevent and treat bacterial-induced infection.

SUMMARY OF THE INVENTION The aim of the present invention is to provide an efficient medicament and method of blocking or inhibiting SSAO or SSAO derived substances such as hydrogen peroxide and thereby preventing and/or treating complications caused by the expression of SSAO or SSAO derived substances such as hydrogen peroxide. In a first aspect the present invention relates to a substance according to claim 1.

In a second aspect the present invention relates to a pharmaceutical composition comprising the substance according to the present invention and pharmaceutically acceptable adjuvants for use as a medicament for blocking or inhibiting semicarbazide sensitive amine oxidase (SSAO) or SSAO derived substances such as hydrogen peroxide, preferably blocking or inhibiting SSAO or SSAO derived substances such as hydrogen peroxide expressed and produced by smooth muscle cells and/or endothelial cells in blood vessels and/or soluble SSAO or SSAO derived substances such as hydrogen peroxide.

In a third aspect the present invention relates to a method blocking or inhibiting SSAO or SSAO derived substances such as hydrogen peroxide preferably expressed by smooth muscle cells and/or endothelial cells in blood vessels and/or soluble SSAO or SSAO derived substances such as hydrogen peroxide in a subject in need thereof comprising administering a first dose or dosage of the substance according to the present invention or administering a first dose or dosage of the pharmaceutical composition according to the present invention.

A fourth aspect of the present invention relates to the use of the substance or the pharmaceutical composition according to the present invention as an immunomodulating agent or an immunomodulating composition.

In a fifth aspect the present invention relates to a pharmaceutical composition comprising the substance according to the present invention and pharmaceutically acceptable adjuvants, wherein the composition further comprises an anaesthetic.

All embodiments disclosed herein are applicable to all aspects unless stated otherwise.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 graph illustrating the effect of the substance of the present invention.

Figure 2, graph illustrating PVAC inhibiting effect on hydrogen peroxide.

Figure 3 photos illustrating PVAC inhibiting effect on IL-8 secretion from primary monocytes. Detection of IL-8 secretion from primary monocytes was achieved by the use of FluoroSpot kit (MabTech, Sweden), a) no stimulation of IL-8 secretion, b) stimulation using R848, c) stimulation using R848 and treated with PVAC.

Figure 4 graph illustrating PVAC inhibiting effect on IL-8 secretion from primary monocytes

Figure 5 graph illustrating PVAC inhibiting effect on the production of IL-6 and IL-8 in primary chondrocytes as determined by ELISA

Figure 6 graphs illustrating PVAC effect on kidney function over time, a) concentration of creatinine, b) concentration of urea, c) concentration of potassium, d) concentration of base excess, and e) concentration of lactate.

ITEMIZED EMBODIMENTS

Item 1. A substance comprising a carrier exhibiting at least one scavenger structure, said scavenger structure comprising a nucleophilic centre complying with the formula

X^l(-R”-)(-R’)_mH_n (formula I) wherein a) X¹ is a single-bonded heteroatom selected amongst N, O and S and exhibits a free electron pair, b) m is 0 or 1 and n is 1 or 2 with the sum of m plus n being 2 for X¹ = N and 1 for X¹ = S and O, c) -R”- is a bivalent organic group providing attachment to the carrier via one of its free valences and direct attachment to the heteroatom X¹ at the other one of its free valences, and d) R’ - is a monovalent organic group directly attached to the heteroatom X¹ via its free valence; for use as a medicament for blocking or inhibiting semicarbazide sensitive amine oxidase (SSAO), preferably blocking or inhibiting SSAO expressed and produced by vascular smooth muscle cells and/or endothelial cells and/or soluble SSAO.

Item 2. The substance according to item 1 wherein the medicament is for blocking or inhibiting SSAO for preventing or treating diabetes related complications.

Item 3. The substance according to item 1 or 2 wherein that the nucleophilic center is or is part of a group selected amongst a) amino groups, preferably primary or secondary amino groups, b) hydrazide groups, a semicarbazide group, a carbazate group, a thiosemicarbazide group, a thiocarbazate group, c) aminooxy groups, and d) thiol groups.

Item 4. The substance according to any of the preceding items, wherein that the carrier is a water-soluble polymer, preferably polyvinyl alcohol.

Item 5. The substance according to any of the preceding items, wherein that the substance is carbazate-functionalized polyvinyl alcohol or semicarbazide functionalized polyvinyl alcohol. Item 6. The substance according to any one of items 1 to 5 wherein the carrier is a polymer and wherein the number of scavengers per monomeric unit of the carrier is 15- 50%, preferably 20-40%.

Item 7. The substance according to any one of items 1 to 5 wherein the carrier is a polymer having a molecular weight of 2.000 dalton or higher, preferably 10.000 dalton or higher, more preferably 10.000 to 30.000 dalton.

Item 8. The substance according to item 2 wherein the diabetes related complication is selected from neuropathy, nephropathy and retinopathy.

Item 9. The substance according to item 1 wherein the blocking or inhibition of SSAO is used for preventing or treating congestive heart failure, neuropathy, nephropathy and retinopathy. Item 10. The substance according to item 1 wherein the blocking or inhibition of SSAO is used for preventing or treating pulmonary inflammation, pulmonary severe acute respiratory syndrome preferably as complication after viral infection including Covid-19 infection and/or acute respiratory distress syndrome (ARDS). Item 11. The substance according to item 1 wherein the blocking or inhibition of SSAO is used for preventing or treating vascular damage, carotid plaque and varicosities.

Item 12. A pharmaceutical composition comprising the substance defined in any of items 1-11 and pharmaceutically acceptable adjuvants for use as a medicament for blocking or inhibiting semicarbazide sensitive amine oxidase (SSAO), preferably blocking or inhibiting SSAO expressed and produced by smooth muscle cells and/or endothelial cells in blood vessels and/or soluble SSAO.

Item 13. The pharmaceutical composition according to item 12 wherein the medicament for blocking or inhibiting SSAO is used for treating diabetes related complications.

Item 14. The composition according to item 12 or 13 wherein the composition is suitable for parenteral administration preferably intravenous administration or subcutaneous administration.

Item 15. The composition according to item 12 or 13 wherein the composition is suitable for oral administration.

Item 16. The composition according to any one of item 12 to 15, wherein the concentration of the substance is 20-500pg/ml, preferably 30-150pg/ml, more preferably 40-120pg/ml.

Item 17. The composition according to any one of item 12 to 15, wherein the concentration of the substance is 20-500pg/mg, preferably 30-150pg/mg, more preferably 40-120pg/mg.

Item 18. The composition according to item 13, wherein the diabetes related complication is selected from neuropathy, nephropathy and retinopathy.

Item 19. The composition according to item 12 wherein the blocking or inhibition of SSAO is used for preventing or treating congestive heart failure, neuropathy, nephropathy and retinopathy.

Item 20. The composition according to item 12 wherein the blocking or inhibition of SSAO is used for preventing or treating pulmonary inflammation, pulmonary severe acute respiratory syndrome (SARS) as complication after viral infection including Covid-19 infection and/or acute respiratory distress syndrome (ARDS).

Item 21. A method of blocking or inhibiting SSAO preferably expressed by smooth muscle cells and/or endothelial cells in blood vessels and/or soluble SSAO in a subject in need thereof comprising administering a first dose or dosage of the substance according to anyone of items 1-9 or administering a first dose or dosage of the pharmaceutical composition according to anyone of items 10-17 to the subject.

Item 22. The method according to item 21 wherein the medicament for blocking or inhibiting semicarbazide sensitive amine (SSAO) is used for preventing or treating diabetes related complications or for preventing or treating congestive heart failure, neuropathy, nephropathy and retinopathy, wherein the diabetes related complication is preferably selected from neuropathy, nephropathy and retinopathy.

Item 23. The method according to item 21 wherein the blocking or inhibition of SSAO is used for preventing or treating vascular damage, carotid plaque, varicosities, neuropathy, nephropathy, retinopathy, pulmonary inflammation or pulmonary severe acute respiratory syndrome (SARS) preferably as complication after viral infection including Covid-19 infection and/or acute respiratory distress syndrome (ARDS).

Item 24. The method according to any one of item 21 to 23, wherein the first dose or dosage is 0.005 to 15mg/kg weight of the subject, preferably 0.05 to 5mg/kg weight of the subject, more preferably 0.5 to 3 mg/kg weight of the subject.

Item 25. The method according to any one of item 21 to 24 wherein the administration is done by parenteral administration preferably intravenous injection, intramuscular or subcutaneous injection.

Item 26. The method according to any one of item 21 to 25 wherein the administration is done by oral administration.

Item 27. The method according to any one of item 21 to 26 wherein the administration is done by inhalation including the use of a nebulizer. Item 28. The method according to item 27 wherein the subject suffers from pulmonary inflammation, pulmonary severe acute respiratory syndrome (SARS) preferably as complication after viral infection including Covid-19 infection and/or acute respiratory distress syndrome (ARDS).

Item 29. The method according to any one of item 21 to 28 wherein the first dose or dosage is administrated 1-3 times/7 days, preferably 1 times/7 days.

Item 30. The method according to any one of item 21 to 28 wherein the subject suffers from acute pulmonary inflammation and wherein the first dose or dosage is administrated 1-3 times/1 day, preferably 1 times/1 day during 1-7 days or preferably until respiratory improvement occurs, where after administrating a first dose or dosage 1-2 times/7 days.

Item 31. The method according to any one of items 18 to 24 wherein the method is or is part of an immunotherapy.

Item 32. Use of the substance according to any one of item 1 to 11 or the composition according to any one of item 12 to 20 as an immunomodulating agent or an immunomodulating composition.

DETAILED DESCRIPTION OF THE INVENTION

The present disclosure is generally directed to substance or composition for used in or methods of preventing and treating complications associated with diabetes and pulmonary inflammation.

Definitions:

"Administration" or "administering", as used herein, refers to providing, contacting, and/or delivering a compound or compounds by any appropriate route to achieve the desired effect. Administration may include, but is not limited to, oral, sublingual, parenteral (e.g., intravenous, subcutaneous, intracutaneous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrathecal, intralesional or intracranial injection), transdermal, topical, buccal, rectal, vaginal, nasal, ophthalmic, via inhalation, and implants. "Co-administered", as used herein, refers to simultaneous or sequential administration of multiple compounds or agents. A first compound or agent may be administered before, concurrently with, or after administration of a second compound or agent. The first compound or agent and the second compound or agent may be simultaneously or sequentially administered on the same day, or may be sequentially administered within 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks or one month of each other. Suitably, compounds or agents are co-administered during the period in which each of the compounds or agents are exerting at least some physiological effect and/or has remaining efficacy.

"Contacting", as used herein as in "contacting a cell," refers to contacting a cell directly or indirectly in vitro, ex vivo, or in vivo (i.e. within a subject, such as a mammal, including humans, mice, rats, rabbits, cats, and dogs). Contacting a cell, which also includes "reacting" a cell, can occur as a result of administration to a subject. Contacting encompasses administration to a cell, tissue, mammal, subject, patient, or human. Further, contacting a cell includes adding an agent to a cell culture. Other suitable methods may include introducing or administering an agent to a cell, tissue, mammal, subject, or patient using appropriate procedures and routes of administration as defined herein. "Effective amount", as used herein, refers to a dosage of compounds or compositions effective for eliciting a desired effect. This term as used herein may also refer to an amount effective at bringing about a desired in vivo effect in an animal, mammal, or human. "Pharmaceutically acceptable", as used herein, pertains to compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject (e.g. human) without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. Each carrier, excipient, etc. must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation.

Subject", as used herein, is intended to include human and non-human animals. In embodiments, the subject is a human. Exemplary human subjects include a human patient having a disorder. The term "non-human animals" includes all vertebrates, e.g., non mammals (such as chickens, amphibians, reptiles) and mammals, such as non-human primates, domesticated and/or agriculturally useful animals (such as sheep, dogs, cats, cows, pigs, etc.), and rodents (such as mice, rats, hamsters, guinea pigs, etc.).

"Treat" or "treating", as used herein, the term a subject having a disorder refers to administering a regimen to the subject, e.g., the administration of a platinum-based therapeutic and/or another agent, such that at least one symptom of the disorder is healed, alleviated, relieved, altered, remedied, ameliorated, or improved. Treating includes administering an amount effective to alleviate, relieve, alter, remedy, ameliorate, improve or affect the disorder or the symptoms of the disorder. The treatment may inhibit deterioration or worsening of a symptom of a disorder.

The present inventors have surprisingly found that administration of the present substance or the present pharmaceutical composition has an inhibitory effect on SSAO or SSAO derived substances such as hydrogen peroxide. As shown in the examples (Example 2) the present substance block or inhibit SSAO or SSAO derived substances such as hydrogen peroxide and thereby the complications caused by the expression of SSAO or SSAO derived substances such as hydrogen peroxide may be prevented, inhibited or treated (Treatment of retinopathy, Example 3), that PVAC has a suppressing effect on the production of IL-8 and IL-6 and prevention of kidney function (Example 4). The present substance has also a treating effect for diabetes related complications such as retinopathy (Example 3). Hence by presenting supporting data (Example 3) in combination with a confirmed mechanism (Inhibition of SSAO or SSAO derived substances, Example 2) it is evident that the present invention relates to a substance, composition and method which may be used to prevent or treat any SSAO or SSAO derived substances related condition or complication.

SSAO is an enzyme found in for example in endothelial cells and smooth muscle cells for example in blood vessels and SSAO or SSAO derived substances such as hydrogen peroxide is believed to be involved in the cause or progression of for example diabetes related complications, congestive heart failure, neuropathy, nephropathy, retinopathy, acute respiratory distress syndrome (ARDS), vascular damage, carotid plaque and varicosities. Also, as compared to smaller agents, the molecular size of the substance according to the present invention effect pharmacokinetic and distribution profiles in a way that may be advantageous for certain indications where a prolonged and directed effect is needed. Other potential benefits include excellent in vivo tolerance and low manufacturing costs. IL-8 is a crucial mediator in the inflammatory cascade. It is acting as a chemokine produced by a variety of cells, including epithelial and endothelial cells. IL-8 exerts a chemotactic effects and induces the migration of neutrophils and monocytes towards a gradient. IL-8 also activates neutrophils. 11-8 is upregulated by several proinflammatory agents, including certain cytokines (e.g. TNF-alpha), oxidative stress (hydrogen peroxide) and high glucose levels. Enhanced levels of IL-8 may facilitate acute and chronic disease in the kidneys (nephropathy), eyes (retinopathy) and nerves (neuropathy), which are typical associated organ failures with type 1 and type 2 diabetes. The serum/plasma level of IL-8 is increased in diabetic patients. PVAC can lower IL-8 production, which may be beneficial for the treatment of nephropathy, retinopathy and neuropathy.

The production of IL-8 by cancer cells is demonstrated in several malignancies, including renal cell carcinoma, malignant melanoma, non-small cell lung cancer and hepatocellular carcinoma. In several cancer patients, the serum/plasma level of both IL-6 and IL-8 is increased. The recruitment of myeloid-derived suppressor cells (MDSC) by IL-8 and the subsequent accumulation in tumors is proposed as an important mechanism, since MDSC suppresses the ability of other immune cells to combat cancer. It is therefore considered as an immune modulating chemokine. IL-8 also counteracts the efficacy of immunotherapies such as anti-CTLA4 (ipilimumab) and anti -PD 1 (pembrolizumab and nivolumab). Thus, the treatment of cancer with PYAC is proposed and may preferably be combined with other immunotherapies for enhanced effect.

First main aspect - A substance for use as a medicament

A substance comprising a carrier which exhibits a plurality of a scavenger structure, said scavenger structure comprising a nucleophilic centre complying with the formula

X¹(-R”-)(-R^,)mH_n (formula I) wherein a) X¹ is a single-bonded heteroatom selected amongst N, O and S and exhibits a free electron pair, b) m is 0 or 1 and n is 1 or 2 with the sum of m plus n being 2 for X¹ = N and 1 for X¹ = S and O, c)-R”-is a bivalent organic group providing attachment to the carrier via one of its free valences and direct attachment to the heteroatom X¹ at the other one of its free valences, and d) R’-is a monovalent organic group directly attached to the heteroatom X¹ via its free valence for use as a medicament for blocking or inhibiting semicarbazide sensitive amine oxidase (SSAO) or SSAO derived substances such as hydrogen peroxide, preferably blocking or inhibiting SSAO or SSAO derived substances such as hydrogen peroxide expressed and produced by vascular smooth muscle cells and endothelial cells and/or soluble SSAO.

The substance blocks or inhibits SSAO or SSAO derived substances such as hydrogen peroxide and thereby prevents or treats any indication or complication related to the expression of SSAO or SSAO derived substances such as hydrogen peroxide. As disclosed above a non-limiting list of such indications or complications are diabetes related complications, neuropathy, nephropathy such as acute kidney failure, retinopathy, pulmonary inflammation such as pneumonia, acute respiratory distress syndrome (ARDS), vascular damage, carotid plaque and varicosities. In one embodiment the substance is used as a medicament for preventing or treating pulmonary severe acute respiratory syndrome preferably as complication after viral infection including Covid-19 infection and/or acute respiratory distress syndrome (ARDS). In another embodiment the substance is used as a medicament for preventing or treating congestive heart failure.

During long time surgery patients may suffer from organ failure or organ damages. In order to reduce such failure or damages the substance may be used as a medicament for preventing complications as a result of surgery such as nephropathy. The substance is then administrated, preferably intravenously, in connection with anaesthesia or is administrated in combination with anaesthetic but preferably prior to the start of the surgery. When administrated in combination with anaesthetic the administration may be done simultaneously or sequentially. The anaesthetic may be any suitable substance administrated in any well-known way such as through inhalation or intravenously.

According to an embodiment of the present substance, either one or both of the organic groups R’-and-R”-, with preference for -R”-, comprise a structure of the formula:

(formula II) wherein a) each of m’, n’ and o’ is 0 or 1, with preference for m’ being 1 with further preference for either one or both of n’ and o’ also being 1, b) each of X² , X³, and X⁴, is selected amongst NH and a heteroatom S or O, with preference for either one or both of X² and X⁴ being selected amongst NH and O with further preference for X³ being selected amongst NH, O and S, c) the left free valence provides binding to a monovalent alkyl group R*-or to the carrier via at least a bivalent alkylene group -R**-, each of which two groups comprises the methylene group-CH2-shown in formula II, and d) the right free valence binds directly to the first heteroatom X¹.

The scavenger structure comprises a first nucleophilic centre which preferably is capable of participating in an addition reaction with the carbonyl group (C=0) of an aldehyde group, or a carbazide group-and/or with a C,C-multiple bond to which one or more electron-withdrawing substituents preferably are directly attached.

Preferred scavenger structures thus have a nucleophilic centre which contain the first heteroatom X¹ together with a structure complying with formula II. According to one embodiment of the present invention, the nucleophilic center is or is part of a group selected amongst: a) amino groups preferably primary or secondary amino groups b) hydrazide groups such as -NH-NH2, e.g. as part of a-CONHMH group, a semicarbazide group such as-NHCONHMH, a carbazate group such as- OCONHNH2, a thiosemicarbazide group such as-NHCSNHMH, a thiocarbazate group such as-OCSNHNH₂ (formation of hydrazone, semicarbazone, thiocarbazone linkages/groups, etc when undergoing addition/elimination reactions with an aldehyde group) c) aminooxy groups, such as-OMB, etc (formation oxime linkages/groups, etc when undergoing addition/elimination reactions with an aldehyde group), d) a thiol group e.g.-SH (Michael addition products are formed when the thiol group reacts with a C,C-double bond. The product may undergo keto-enol tautomerisation when the double bond is a,b to a keto-or aldehyde-carbonyl, see above).

These scavenger structures are believed to be effective when it comes to binding or interact with SSAO or SSAO derived substances such as hydrogen peroxide and thereby blocking or inhibiting SSAO or SSAO derived substances such as hydrogen peroxide.

According to a still further embodiment of the substance, the carrier is a macromolecular carrier and/or is water-soluble or water-insoluble and preferably exhibits polymer structure. The carrier may be water-insoluble and define a support and/or the substance is attached to a water-insoluble support. In one embodiment the carrier is a water-soluble polymer, preferably polyvinyl alcohol, polysaccharides preferably glucose amino glucan, hyaluronic acid or dextran. In a preferred embodiment the carrier is a water-soluble polymer preferably polyvinyl alcohol. By using a water soluble carrier the substance is easier dissolved making it easier to prepare and it is also believed to increase the efficiency of the substance when administered. Polyvinyl alcohol makes it also possible to provide several scavenger structures per carrier.

According to another embodiment, the substance has a scavenger structure capable of undergoing an addition reaction with a carbonyl group of an aldehyde group, or a carbazide group and/or with a carbon-carbon multiple bond to which is directly attached a carbonyl group, such as a semicarbazide group or an aldehyde group.

In one embodiment, the substance is carbazate-functionalized polyvinyl alcohol or a semicarbazide-functionalized polyvinyl alcohol. The substance is preferably soluble in aqueous liquids such as water, body fluids, such as blood, serum, plasma, urine, lymph, lachrymal fluid, intestinal juice, gastric juice, saliva, synovial fluid, etc. The selection of suitable carriers depends on the requirements of a particular use. The typical carrier is selected amongst macromolecular compounds, i.e. is a compound which has a molecular weight of > 2000 dalton, preferably > 10000 dalton or > 50000 dalton, and preferably exhibits a polymeric structure, i.e. is a polymer which may be a homopolymer, copolymer or a chemical adduct between two or more polymers of different polymeric structure. In a preferred embodiment the carrier has a molecular weight of 10.000-30. OOOdalton. Other suitable carriers may have molecular weights < 2000 dalton and exhibit polymeric structure as indicated by the possibility of the low numbers of monomeric units discussed below, e.g. > 20 and < 100. The term “adduct polymer” in this context means a product formed by reacting two polymers exhibiting mutually groups capable of forming covalent bonds that link the two polymers together upon reaction of the two mutually groups with each other. See for instance WO 2009108100 (IPR-Systems AB) and references cited therein. Suitable macromolecular carriers may thus be selected amongst synthetic polymers (= man-made polymers), biopolymers (nature-made polymers such as polysaccharides, polypeptides, proteins, etc) and biosynthetic polymers where “biosynthetic polymer” refers to a macromolecular carrier or compound exhibiting both a synthetic polymeric structure and a biopolymeric structure.

The scavenger structure including the first, the optional second nucleophilic centre and the various heteroatoms discussed for the scavenger structures are typically part of one and the same organic group/substituent attached to the macromolecular carrier. In certain variants different parts of a scavenger structure may be part of different group s/substituents attached to the carrier and/or part of the carrier. Sizes/molecular weights of suitable carrier polymers will among others depend on the actual application/use of the composition/method of the invention. Thus suitable polymeric carriers with respect to a particular polymeric structure and/or size may vary within a wide interval. Thus as a rule the number of monomeric subunits (mean value) of a polymer present in the carrier may be > 20 or > 100 or > 200 or > 300 or > 500 or >

1000 or > 2000 or > 20000 or > 50 000 and/or < 50000 or < 20000 or < 2000 or < 1000 or < 500 or < 300 or < 200, or < 100 (with the proviso that >-limit always is lower than a <-limit when these values are combined to define intervals). Preferred numbers of monomeric units may in some cases be found in the interval of 200 - 600. In a preferred embodiment the carrier is a water soluble polymer, preferably polyvinyl alcohol, having a molecular weight of 10.000-30. OOOdalton.

Suitable numbers of scavenger structures or nucleophilic centres per monomeric unit of a polymer of the carrier will also depend on the use, the scavenger structure, etc, and may thus be found within a wide interval, such as < 80%, preferably < 50%, <40% or < 30% with lower limits being preferably 0.01% or 0.1% or 1% or 5% or 10% or 15% where 100% corresponds to one scavenger structure or nucleophilic centre per monomeric unit. For scavenger structures containing two or more nucleophilic centres the number of nucleophilic centres per monomeric unit may exceed 100%, such as > 100% or > 125% or > 150%. In a preferred embodiment the number of scavenger structures or monomeric unit is 15-50%, more preferably 20-40%.

The substance according to the present invention may be synthesized according to well- known protocols, for instance of the kinds given in WO 2009108100 (IPR-Systems AB) and references cited therein which are hereby incorporated by reference.

All variants, embodiments and examples of the first main aspect can be combined with the second and third main aspects unless expressly stated otherwise.

Second and fifth main aspect - Pharmaceutical composition

According to a second aspect, there is provided a pharmaceutical composition for use as a medicament in the prevention or treatment of diabetes related conditions or for use as a medicament for blocking or inhibiting SSAO or SSAO derived substances such as hydrogen peroxide, comprising the substance according to the present invention and pharmaceutically acceptable adjuvants. According the fifth aspect there is a pharmaceutical composition comprising the substance as defined above and pharmaceutically acceptable adjuvants, wherein the composition further comprises an anaesthetic.

The pharmaceutical formulation or composition according to the present invention may conveniently be presented in unit dosage form and may be prepared by any methods known in the art of pharmacy. Such methods include the step of bringing into association the active compound(s) or substance with the carrier which constitutes one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association the active compound or substance with liquid carriers or finely divided solid carriers or both, and then if necessary, shaping the product.

The pharmaceutical composition may be in the form of liquids, solutions, suspensions, emulsions, elixirs, syrups, tablets, lozenges, granules, powders, capsules, cachets, pills, ampoules, suppositories, pessaries, ointments, gels, pastes, creams, sprays, mists, foams, lotions, oils, boluses, electuaries, or aerosols.

Pharmaceutical composition suitable for oral administration (e.g. by ingestion) may be presented as discrete units such as capsules, cachets or tablets, each containing a predetermined amount of the active compound or substance; as a powder or granules; as a solution or suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion; as a bolus; as an electuary; or as a paste.

A tablet may be made by conventional means, e.g., compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared by compressing in a suitable machine the active compound or substance in a free-flowing form such as a powder or granules, optionally mixed with one or more binders (e.g. povidone, gelatin, acacia, sorbitol, tragacanth, hydroxypropylmethyl cellulose); fillers or diluents (e.g. lactose, microcrystalline cellulose, calcium hydrogen phosphate); lubricants (e.g. magnesium stearate, talc, silica); disintegrants (e.g. sodium starch glycolate, cross- linked povidone, cross-linked sodium carboxymethyl cellulose); surface-active or dispersing or wetting agents (e.g. sodium lauryl sulfate); and preservatives (e.g. methyl p- hydroxybenzoate, propyl p-hydroxybenzoate, sorbic acid). Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent. The tablets may optionally be coated or scored and may be ftod so as to provide slow or controlled release of the active compound or substance therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile. Tablets may optionally be provided with an enteric coating, to provide release in parts of the gut other than the stomach.

Pharmaceutical composition suitable for parenteral administration (e.g. by injection, including cutaneous, subcutaneous, intramuscular, intravenous and intradermal), include aqueous and nonaqueous isotonic, pyrogen-free, sterile injection solutions which may contain antioxidants, buffers, preservatives, stabilizers, bacteriostats, and solutes which render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents, and liposomes or other microparticulate systems which are designed to target the compound to blood components or one or more organs. Examples of suitable isotonic vehicles for use in such formulations include Sodium Chloride Injection, Ringer's Solution, or Lactated Ringer's Injection. The formulations may be presented in unit-dose or multi-dose sealed containers, for example, ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, and tablets. Formulations may be in the form of liposomes or other microparticulate systems which are designed to target the active compound or substance to blood components or one or more organs.

In one embodiment the composition is suitable intravenous administration and wherein the composition further comprises an anaesthetic. The dose of the substance may be 1- 100pm/ml serum, preferably 2-50pg/ml serum, more preferably 3-1 Opg/ml serum. The anaesthetic is preferably an opioid or a non-opioid.

Pharmaceutical composition suitable for topical administration (e.g. transdermal, intranasal, ocular, buccal, and sublingual) is preferably formulated as an ointment, cream, suspension, lotion, powder, solution, past, gel, spray, aerosol, or oil. Alternatively, a formulation may comprise a patch or a dressing such as a bandage or adhesive plaster impregnated with active compounds or substance and optionally one or more excipients or diluents.

Pharmaceutical composition suitable for topical administration in the mouth include lozenges comprising the active compound or substance in a flavored basis, usually sucrose and acacia or tragacanth; pastilles comprising the active compound or substance in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active compound in a suitable liquid carrier.

Pharmaceutical composition suitable for topical administration to the eye also include eye drops wherein the active compound or substance is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active compound or substance.

Pharmaceutical composition suitable for nasal administration, wherein the carrier is a solid, include a coarse powder having a particle size, for example, in the range of about 20 to about 500 microns which is administered in the manner in which snuff is taken, i.e. by rapid inhalation through the nasal passage from a container of the powder held close up to the nose. Suitable formulations wherein the carrier is a liquid for administration as, for example, nasal spray, nasal drops, or by aerosol administration by nebulizer, include aqueous or oily solutions of the active compound or substance.

Pharmaceutical composition suitable for administration by inhalation include those presented as an aerosol spray from a pressurized pack, with the use of a suitable propellant, such as dichlorodifluoromethane, trichlorofluoromethane, dichoro- tetrafluoroethane, carbon dioxide, or other suitable gases. Further formulations suitable for inhalation include those presented as a nebulizer. In one embodiment the composition is suitable for administration done by inhalation including the use of a nebulizer.

Pharmaceutical composition suitable for topical administration via the skin include ointments, creams, and emulsions. When formulated in an ointment, the active compound or substance may optionally be employed with either a paraffinic or a water-miscible ointment base.

Alternatively, the active compound or substances may be formulated in a cream with an oil-in-water cream base. If desired, the aqueous phase of the cream base may include, for example, at least about 30% w/w of a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups such as propylene glycol, butane-1, 3 -diol, mannitol, sorbitol, glycerol and polyethylene glycol and mixtures thereof. The topical formulations may desirably include a compound which enhances absorption or penetration of the active compound or substance through the skin or other affected areas. Examples of such dermal penetration enhancers include dimethylsulfoxide and related analogues.

When formulated as a topical emulsion, the oily phase may optionally comprise merely an emulsifier (otherwise known as an emulgent), or it may comprise a mixture of at least one emulsifier with a fat or an oil or with both a fat and an oil. Preferably, a hydrophilic emulsifier is included together with a lipophilic emulsifier which acts as a stabilizer. It is also preferred to include both an oil and a fat. Together, the emulsifier(s) with or without stabilizer(s) make up the so-called emulsifying wax, and the wax together with the oil and/or fat make up the so-called emulsifying ointment base which forms the oily dispersed phase of the cream formulations.

Suitable emulgents and emulsion stabilizers include Tween 60, Span 80, cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodium lauryl sulfate. The choice of suitable oils or fats for the formulation is based on achieving the desired cosmetic properties, since the solubility of the active compound or substance in most oils likely to be used in pharmaceutical emulsion formulations may be very low. Thus, the cream should preferably be a non-greasy, non-staining and washable product with suitable consistency to avoid leakage from tubes or other containers. Straight or branched chain, mono-or dibasic alkyl esters such as diisoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branched chain esters known as Crodamol CAP may be used, the last three being preferred esters. These may be used alone or in combination depending on the properties required. Alternatively, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils can be used.

Pharmaceutical composition suitable for rectal administration may be presented as a suppository with a suitable base comprising, for example, cocoa butter or a salicylate. Pharmaceutical composition suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active compound or substance, such carriers as are known in the art to be appropriate.

In one embodiment the substance is as a pharmaceutical composition in which the substance is: a) in dry form, for instance as free particles, b) in dissolved form, typically in an aqueous liquid medium, or c) in suspended/dispersed form, i.e. as water-insoluble particles suspended in an aqueous liquid medium.

The term “dissolved” in this context means that the substance is present as a solute. The substance particles comprise substance in a pure form or diluted with some solid material. Useful concentrations of the substance in formulations according to (b) can be found within a broad interval.

The composition may in addition to the substance contain buffers, salts, etc required for enabling acceptable conditions in vivo for the patient and for the reaction of the substance. These constituents may be co-formulated with the substance in pharmaceutical composition.

In one embodiment the pharmaceutical composition is suitable for parenteral administration, preferably for intravenous administration.

In one embodiment the concentration of the substance in the pharmaceutical composition according to the present invention is 20-2500pg/ml, preferably 100-2000 pg/ml, more preferably 500-2000 pg/ml. In another embodiment the concentration is 20-500pg/ml, preferably 30-150pg/ml, more preferably 40-120pg/ml.

The amount of the dose is preferably expressed as pg/ml serum. In one embodiment the dose may be 1-lOOpm/ml serum, preferably 2-50pg/ml serum, more preferably 3- 10pg/ml serum.

All variants, embodiments and examples of the second main aspect can be combined with the first and third main aspects unless expressly stated otherwise.

Third main aspect - Method of blocking or inhibiting semicarbazide sensitive amine oxidase (SSAO) or SSAO derived substances

This aspect relates to a method of blocking or inhibiting SSAO or SSAO derived substances such as hydrogen peroxide by using the present substance or pharmaceutical composition (defined above under the first and second main aspects) which may be done for prevention or treatment of diabetes related complications, congestive heart failure, pulmonary inflammation, vascular damage, carotid plaque and/or varicosities.

The substance or the pharmaceutical composition disclosed herein is administered to a subject or a patient in need thereof. The patient may be an animal or a human. The administration ways or routes may vary according to the specific indication or medical situation and are part of the knowledge of a medical practitioner.

The administration of the first dose may be done locally or systemically or in combination. Preferred administration of the first dose is as disclosed above. Considering that SSAO is found in the blood vessels the preferred administration is intravenous administration. For the treatment of diabetes related conditions a preferred way of administration is subcutaneous since that is a common administration of insulin. In a preferred embodiment the administration is done by parenteral administration preferably intravenous injection, intramuscular or subcutaneous injection. Example 3 shows the effect of improving the eyesight after parenteral administration (subcutaneous) of the present substance. In another preferred embodiment the administration is done by oral administration.

In another preferred embodiment the administration is done by inhalation including the use of a nebulizer. This is specifically preferred for subjects suffering from pulmonary inflammation, pulmonary severe acute respiratory syndrome (SARS) preferably as complication after viral infection including Covid-19 infection and/or acute respiratory distress syndrome (ARDS). In one embodiment the subjects is a diabetes patient and the present composition may then be administrated 1-2 times/week subcutaneously in order to prevent or treat the progression of nephrophaty, neurophaty and/or retinopathy.

In general, a suitable dose of the substance is in the range of about 0. lmg to about 1.5mg per kilogram body weight of the subject per administration. In one embodiment the first dose or dosage is 0.05 to 5mg/kg weight of the subject, more preferably 0.5 to 3 mg/kg weight of the subject or preferably O.lmg to about 1.5mg/kg weight of the subject. In another embodiment the first dose or dosage is administrated 1-3 times/7 days, preferably 1-2 times/7 days, more preferably 1 times/7days. A second dose or dosage is preferably 0.005 to 15mg/kg weight of the subject, preferably 0.05 to 5mg/kg weight of the subject, more preferably 0.1 to 1.5 mg/kg weight of the subject. In one embodiment the second dose or dosage is the same as the first dose or dosage. The second dose or dosage is administrated after a first treatment time of the first dose or dosage. The first treatment time is preferably 7 days or longer, or 14 days or longer.

The amount of the dose is preferably expressed as pg/ml serum. In one embodiment the dose may be 1-100pm/ml serum, preferably 2-50 pg/ l serum, more preferably 3- 10pg/ml serum. As seen in Example 2 the present substance (PVAC) was highly efficient and doses below 20 ug/ml serum are predicted to be suitable in an vivo situation which corresponds to a dose of 60 mg in adult patient.

In one embodiment the subject suffers from acute pulmonary inflammation and wherein the first dose or dosage is administrated 1-3 times/ 1 day, preferably 1 times/ 1 day during 1-7 days or preferably until respiratory improvement occurs, where after a first dose or dosage is administrated 1-2 times/7 days. In one embodiment the substance or the composition is used in the prevention or treatment of congestive heart failure, nephropathy, neuphropaty and/or retinopathy and the first dose or dosage is 0.1 to 15mg/kg weight of the subject, preferably 0.15 to 5mg/kg, more preferably 0.1 to 1.5mg/kg. The substance is preferably administrated intra venously. When the substance or the composition is used for treating or preventing congestive heart failure a second dose or dosage may be administrated as disclosed above. The second dose or dosage is preferably 0.005 to 15mg/kg weight of the subject, preferably 0.05 to 5mg/kg weight of the subject, more preferably 0.1-1.5 mg/kg weight of the subject. In one embodiment the second dose or dosage is the same as the first dose or dosage. The second dose or dosage is administrated after a first treatment time of the first dose or dosage. The first treatment time is preferably 7 days or longer, or 14 days or longer.

In order to reduce such organ failure or organ damages during long time surgery the substance or the pharmaceutical composition may be used as a medicament for preventing complications as a result of surgery such as nephropathy. The substance is then administrated, preferably intravenously, in connection with anaesthesia or is administrated in combination with anaesthetic but preferably prior to the start of the surgery. When administrated in combination with anaesthetic the administration may be done simultaneously or sequentially. The anaesthetic may be any suitable substance administrated in any well-known way such as through inhalation or intravenously. In one embodiment the pharmaceutical composition is suitable intravenous administration and wherein the composition further comprises an anaesthetic. The dose of the substance may be 1-100pm/ml serum, preferably 2-50pg/ml serum, more preferably 3-10pg/ml serum. The anaesthetic is preferably an opioid or a non-opioid.

The substance blocks or inhibits SSAO and thereby prevents or treats any indication or complication related to the expression of SSAO or SSAO derived substances. As disclosed above a non-limiting list of such indications or complications are diabetes related complications, neuropathy, nephropathy, retinopathy, pulmonary inflammation such as pneumonia, acute respiratory distress syndrome (ARDS), vascular damage, carotid plaque and/or varicosities. In one embodiment the method is for preventing or treating pulmonary severe acute respiratory syndrome preferably as complication after viral infection including Covid-19 infection and/or acute respiratory distress syndrome (ARDS). In one embodiment the method is or is part of an immunotherapy. In another embodiment the method is for preventing or treating congestive heart failure.

All variants, embodiments and examples of the third main aspect can be combined with the first and second main aspects unless expressly stated otherwise.

Fourth main aspect - Use as an immunomodulating agent or composition

As described above the present substance or pharmaceutical composition may be used as an immunomodulating agent or composition.

This may be done by administrating the substance or composition as disclosed herein. Preferably the use is done by administrating a first dose locally or systemically or in combination. Preferred administration of the first dose is as disclosed above. Considering that SSAO is found in the blood vessels the preferred administration is intravenous administration. For the treatment of diabetes related conditions a preferred way of administration is subcutaneous since that is a common administration of insulin.

In one embodiment the first dose is 0.005 to 15mg/kg weight of the subject, preferably 0.05 to 5mg/kg weight of the subject, more preferably 0.1 to 1.5 mg/kg weight of the subject. In another embodiment the administration is done 2-3 times/7 days, preferably 2 times/7 days.

The substance blocks or inhibits SSAO or SSAO derived substances such as hydrogen peroxide and thereby prevents or treats any indication or complication related to the expression of SSAO or SSAO derived substances such as hydrogen peroxide. As disclosed above a non-limiting list of such indications or complications diabetes related complications, neuropathy, nephropathy, retinopathy, pulmonary inflammation such as pneumonia, acute respiratory distress syndrome (ARDS), vascular damage, carotid plaque and/or varicosities.

All variants, embodiments and examples of the third main aspect can be combined with the first and second main aspects unless expressly stated otherwise. Dosages

It will be appreciated that appropriate doses and dosages of the active compound or substances, and compositions comprising the active compound or substances, can vary from patient to patient. All dosages disclosed here below are applicable to all aspects disclosed herein.1

Determining the optimal dosage will generally involve the balancing of the level of therapeutic benefit against any risk or deleterious side effects of the treatments described herein. The selected dosage level will depend on a variety of factors including, but not limited to, the activity of the particular compound, the route of administration, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds, and/or materials used in combination, and the age, sex, weight, condition, general health, and prior medical history of the patient. The amount of compound and route of administration will ultimately be at the discretion of the physician, although generally the dosage will be to achieve local concentrations at the site of action which achieve the desired effect without causing substantial harmful or deleterious side-effects. Administration in vivo can be affected in one dose, continuously or intermittently (e.g. in divided doses at appropriate intervals) throughout the course of treatment. Methods of determining the most effective means and dosage of administration are well known to those of skill in the art and will vary with the formulation used for therapy, the purpose of the therapy, the target cell being treated, and the subject being treated. Single or multiple administrations can be carried out with the dose level and pattern being selected by the treating physician.

A suitable dose or dosage of the substance is in the range of about 0.005-15mg/kg weight of the subject, preferably 0.05 to 5mg/kg weight of the subject, more preferably 0.1 to 1.5 mg/kg weight of the subject. During a course of a week or seven days an amount of 50- 140mg may be administered to the subject. Preferably the dosing or administration should be done 2-3 times/7days, preferably 1-2 times/7 days in order to avoid accumulation of the substance in the subject. When formulating the pharmaceutical compositions described herein, the clinician may utilize preferred dosages as warranted by the condition of the subject being treated. For example, in one embodiment, the substance according to the invention may be administered at a dosing schedule described herein, e.g., once every one, two, three, four, five, six or seven days.

Also, in general, the substance according to the invention and an optional additional chemotherapeutic agent(s) or other agents do not have to be administered in the same pharmaceutical composition, and may, because of different physical and chemical characteristics, have to be administered by different routes. The determination of the mode of administration and the advisability of administration, where possible, in the same pharmaceutical composition, is well within the knowledge of the skilled clinician. The initial administration can be made according to established protocols known in the art, and then, based upon the observed effects, the dosage, modes of administration and times of administration can be modified by the skilled clinician.

The actual dosages of the compounds employed may be varied depending upon the requirements of the subject and the severity of the condition being treated. Determination of the proper dosage for a particular situation is within the skill of the art. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small amounts until the optimum effect under the circumstances is reached. Diabetes related complications

The substance and the composition according to the present invention may be used to prevent or treat complications caused by or related to diabetes.

Insulin tend to increase the expression of SSAO which is believed to be one of the reasons for some of the complications seen in diabetes patients. Without being bound by theory the substance according to the present invention is believed to bind to and thereby block or inhibit SSAO which then prevents, limits or treats the complication relating to the expression of SSAO or SSAO derived substances such as hydrogen peroxide. A non-limiting list of diabetes related complications that can be prevented or treated are vascular damage, neuropathy, nephropathy and/or retinopathy. EXPERIMENTAL PART

EXAMPLE 1 - SYNTHESIS OF CARBAZATE-FUNCTIONALIZED POLYVINYL ALCOHOL (PVAC)

Polyvinyl alcohol (5 g, 13-23 kDa) was dissolved in dimethyl sulfoxide (100 mL) while stirring for 1 hour at 80°C under argon gas. Carbonyl diimidazole (10 g) was added and stirring continued at room temperature overnight. Hydrazine hydrate (10 mL) was then added, the reaction stirred overnight, and the product collected and purified by repeated precipitation in ethanol. The degree of substitution was determined spectrophotometrically by performing a trinitrobenzene sulfonic acid assay described elsewhere (Stephen L. Snyder and Philip Z. Sobocinski; Analytical Biochemistry 64, 284- 288, 1975).

EXAMPLE 2 - INHIBITION OF SSAO OR SSAO DERIVED SUBSTANCES SUCH AS

HYDROGEN PEROXIDE

A study performed by the use of Fluoro:SSAO kit (Cell Technology Inc. USA) was conducted according to instructions. Two different doses of PVAC were applied; 20 ug/ml and 80 ug/ml. The inhibitory effect, as assessed by the measurement of hydrogen peroxide-conjugated chromophore, was dose dependent showing an approximately 80 % reduction of enzyme activity with 20 ug/ml and 90 % reduction with 80 ug/mL PVAC. Thus, the PVAC effect was highly efficient and doses below 20 ug/ml serum are predicted to be suitable in an in vivo situation (corresponding to a dose of 60 mg in adult patient).

A luminesce assay was performed to assess if PVAC exerts effect on hydrogen peroxide- induced luminescence. White solid Tecan 96 well plate were used and read within 5 minutes after mixing (see details below). Figure 2.

Conclusion: PVAC diminishes hydrogen peroxide induced luminescence. This implies that PVAC reduces oxidative stress through quenching hydrogen peroxide. Detailed materials and methods:

Method: HRP- HiC -Luminol assay.

Materials: HRP: Thermo Fischer Cat nr. 31490

Luminol- H2O2 (ECL) Thermo Fischer Cat nr 321091.

PVAC. Specific polymers Batch SP-POL-114 HRP: 1 mg/ml in dd water. Use 10 ul per assay

ECL. 50/50 mix of component A (luminol) and B (H2O2). Use 200 ul per assay PVAC: 250 ug/ml (low) or 2500 ug/ml (high) in dd water. Use 200 ul per assay.

DD Water was used as control. 200 ul per assay.

EXAMPLE 3 - TREATMENT OF RETINOPATHY

A single subcutaneous PVAC injection of 2 mg PVAC resulted in improved clear eye vision in human subject with body weight 80 kg. The effect was apparent after a few days and lasted for 1-2 weeks.

EXAMPLE 4 - PREVENTION OR TREATMENT OF NEPHROPATHY A study was performed to study the effect of PVAC on IL-8 and IL-6 expression.

PVAC was shown to reduce the induction of IL-8 expression in monocytes at concentration 5 ug/mL, which is within the serum level range of PVAC after therapeutic administration and is therefore highly relevant from a interventional and treatment perspective, Figure 3 and 4. IL-8 is a chemokine with chemotactic and activating effects on white blood cells, including neutrophils. Increased IL-8 levels in serum or plasma is shown to be associated with several diseases, including nephropaty, retinopathy and neuropathy in human subjects. The detection of IL-8 secretion from primary monocytes was achieved by the use of

FluoroSpot kit (MabTech, Sweden). The assay allows for detection and quantification of IL-8 producing cells. The cells were incubated with PVAC (5ug/mL) for 30 minutes prior subsequent treatment with TLR7/8 agonist R848. In this assay, PVAC reduces the number of IL-8 producing cells by approximately 50 %.

PVAC inhibits the production of IL-6 and IL-8 in primary chondrocytes as determined by ELISA. High Mobility Group Box-1 (HMGB-1, TLR2/TLR4/RAGE ligand) was pre incubated with PVAC or PBS (control) prior treatment of cells. PVAC at concentration range 0.8-100 ug/mL reduced IL-6 and IL-8 production induced by HMGB-1 (but not LPS) by approximately 50 %. Figure 5.

A study was performed to test how the function of the kidney was affected during surgery.

Twelve male Wistar Han rats were included in the study. The animals were divided into four groups of three, to have the test item administered in doses of 0.15 (LD, low dose), 1.5 (MD, medium dose) or 15 mg/kg (HD, high dose) or the vehicle (NaCl) administered once intravenously during the initiation of anesthesia.

Animals were anaesthetised with urethane after which a tracheotomy was performed to ensure the animal was sufficiently ventilated. Thereafter, an indwelling catheter was placed into a lateral vein for fluid replacement by infusion via an external pump, or test item/vehicle administration by intravenous bolus injection. The urinary bladder was catheterised to allow drainage of urine. An indwelling catheter was placed in the carotid artery to monitor blood pressure changes, and for blood sampling purposes.

Blood was collected at baseline and once an hour following test item administration for assessment of Na+, K+, C1-, iCa2+, TC02, glucose, blood urea nitrogen, creatinine, haematocrit, haemoglobin, anion gap, lactate, pC02, p02, s02, pH, BEecf and HC03. Animals were monitored up to 5 hr after administration of test item.

Outcome: Animals in the vehicle group demonstrated marked alterations in blood parameters at the final measurement, with increases in anion gap, lactate, creatinine, potassium and urea, as well as decreases in glucose, HC03, pC02, TC02, and BEecf. These changes were not evident in any test-item treated groups (PVAC 0.15 mg/kg, 1.5 mg/kg and 15 mg/kg). These results imply that PVAC protects the kidneys from injury during surgery and the development of metabolic acidosis. Figure 6.

Claims

C L A I M S

1. A substance comprising a carrier exhibiting at least one scavenger structure, said scavenger structure comprising a nucleophilic centre complying with the formula

X¹(-R”-)(-R^,)mH_n (formula I) wherein a) X¹ is a single-bonded heteroatom selected amongst N, O and S and exhibits a free electron pair, b) m is 0 or 1 and n is 1 or 2 with the sum of m plus n being 2 for X¹ = N and 1 for X¹ = S and O, c) -R”- is a bivalent organic group providing attachment to the carrier via one of its free valences and direct attachment to the heteroatom X¹ at the other one of its free valences, and d) R’ - is a monovalent organic group directly attached to the heteroatom X¹ via its free valence; wherein the nucleophilic center is or is part of a group selected amongst hydrazide groups, a semicarbazide group, a carbazate group, a thiosemicarbazide group, a thiocarbazate group, and wherein the carrier is a water-soluble polymer, preferably polyvinyl alcohol, polysaccharides preferably glucose amino glucan, hyaluronic acid or dextran; for use as a medicament for preventing or treating congestive heart failure, neuropathy, nephropathy and/or retinopathy.

2. The substance for use according to claim 1, wherein that the carrier is a water-soluble polymer, preferably polyvinyl alcohol.

3. The substance for use according to claim any of the preceding claims, wherein that the substance is carbazate-functionalized polyvinyl alcohol or semicarbazide functionalized polyvinyl alcohol.

4. The substance for use according to any one of claims 1 to 3 wherein the carrier is a polymer and wherein the number of scavengers per monomeric unit of the carrier is 15-50%, preferably 20-40%.

5. The substance for use according to any one of claims 1 to 4 wherein the carrier is a polymer having an average molecular weight of 2.000 dalton or higher, preferably 10.000 dalton or higher, more preferably 10.000 to 30.000 dalton.

6. The substance for use according to claim 1 to 5 wherein the substance is administrated in connection with anaesthesia or is administrated in combination with anaesthetic.

7. A pharmaceutical composition comprising the substance defined in any of claims 1-6 and pharmaceutically acceptable adjuvants for use as a medicament for blocking or inhibiting for preventing or treating congestive heart failure, neuropathy, nephropathy and/or retinopathy.

The pharmaceutical composition for use according to claim 7 wherein the composition is suitable for parenteral administration preferably intravenous administration or subcutaneous administration.

9. The pharmaceutical composition for use according to claim 7 or 8 wherein the composition is suitable for oral administration.

10. The pharmaceutical composition for use according to any one of claim 7 to 9, wherein the concentration of the substance is 100-2500pg/ml

11. The pharmaceutical composition for use according to claim 8 wherein the composition is suitable intravenous administration and wherein the composition further comprises an anaesthetic.

12. A pharmaceutical composition comprising the substance defined in any of claims 1- 6 and pharmaceutically acceptable adjuvants, wherein the composition further comprises an anaesthetic.

13. The pharmaceutical composition according to claim 12 wherein the composition is suitable intravenous administration wherein the concentration of the substance preferably is 100-2500pg/ml.

14. A method of preventing or treating congestive heart failure, neuropathy, nephropathy and/or retinopathy in a subject in need thereof comprising administering a first dose or dosage of the substance according to anyone of claims 1-6 or administering a first dose or dosage of the pharmaceutical composition according to anyone of claims 7- 11 to the subject.

15. The method according to any one of claim 12, wherein the first dose or dosage is 0.05 to 5mg/kg weight of the subject, preferably 0.1 to 1.5mg/kg weight of the subject.

16. The method according to any one of claim 12 to 13 wherein the administration is done by parenteral administration preferably intravenous injection, intramuscular or subcutaneous injection.

17. The method according to claim 12 wherein the administration is done to result in a substance concentration of 1-100pg/ml serum, preferably 1-10pg/ml serum, more preferably 3-6pg/ml serum.

18. The method according to any one of claim 12 to 15 wherein the administration is done by oral administration.

19. The method according to any one of claim 12 to 16 wherein the administration is done by inhalation including the use of a nebulizer.

20. The method according to any one of claim 12 to 19 wherein the first dose or dosage is administrated 1-3 times/7 days, preferably 1 times/7 days.

21. The method according to any one of claim 12 to 20 wherein the subject suffers from acute pulmonary inflammation and wherein the first dose or dosage is administrated 1-3 times/1 day, preferably 1 times/1 day during 1-7 days or preferably until respiratory improvement occurs, where after administrating a first dose or dosage 1-2 times/7 days.

22. The method according to any one of claims 12 to 21 wherein the method is or is part of an immunotherapy.

23. The method according to claim 12 wherein a first dose or dosage is administrated in connection with anaesthesia or is administrated in combination with an anaesthetic.

24. Use of the substance according to any one of claim 1 to 6 or the composition according to any one of claim 7 to 11 as an immunomodulating agent or an immunomodulating composition.