Inositol phosphate compounds for use in increasing tissular perfusion

Abstract

To provide: compounds for use in increasing the tissular perfusion and / or oxygen supply in a subject requiring the increasing, particularly in peripheral arterial disease; and pharmaceutical compositions comprising the compounds.SOLUTION: The invention provides compounds of general formula I for use in increasing the tissular perfusion and / or oxygen supply in a subject requiring the increasing. [In the formula, R1, R3, R5, R7, R9 and R11 are independently selected from among OH, -O-P(O)(OH)2, -O-S(O)(OH)2 and -O-P(S)(OH)2 groups and heterologous moieties.SELECTED DRAWING: None

Description

[Technical field]

[0001]

[0001] This application claims the benefit of priority to EP19382061.0, filed January 30, 2019, and US62 / 913,259, filed October 10, 2019.

[0002] The present invention relates to the use of inositol phosphates (IPs), their analogs and derivatives to increase tissue perfusion and / or oxygenation. The present invention also relates to pharmaceutical compositions comprising said IPs, their analogs and derivatives and their use in animal and human health. [Background technology]

[0002]

[0003] Peripheral arterial disease (PAD) is a common disorder characterized by narrowing and / or blockage of the lower limb arteries, resulting in reduced muscle perfusion and oxygen delivery. PAD represents a major public health problem and carries a high risk of long-term suffering. PAD increases the risk of tissue death (gangrene), amputation, and premature death.

[0003]

[0004] PAD is the result of ischemia in the lower limbs. Its main cause is atherosclerosis. In its mild form, PAD may be limited to intermittent claudication and pain in the lower limbs. Lower limb PAD is the main cause of disability and loss of mobility in elderly men and women, and has a decisive impact on quality of life.

[0004]

[0005] The prevalence of PAD is approximately 12% in the adult population. This prevalence increases to over 20% in the population over 70 years of age. PAD currently affects over 8 million men and women in the United States alone. It is estimated that over 200 million individuals worldwide suffer from PAD. The prevalence of PAD is likely to increase in the near future as the general population ages and the incidence of obesity-related type 2 diabetes increases. Cigarette smoking is another significant risk factor. Patients with PAD experience increased cardiovascular (CV) morbidity and mortality, faster rates of functional decline, and greater rates of loss of mobility compared to the general population.

[0005]

[0006] The goals of protocols for the treatment of patients with PAD include reducing CV event rates, improving functional activity, and preventing functional decline and loss of mobility. Restoring or improving blood perfusion to the limbs can help achieve these goals.

[0006]

[0007] While endovascular and lower extremity revascularization procedures significantly improve walking ability in PAD patients, revascularization procedures are not a treatment option for many patients because of the presence of coexisting disease or because the location and type of arteriosclerotic disease in the lower extremities is not amenable to revascularization. Revascularization procedures are invasive, costly, and carry risks, especially for elderly patients. For these reasons, there is a demand from clinicians for effective, achievable, and tolerable medical therapies that improve lower extremity function in PAD patients.

[0007]

[0008] Currently, only two medications have been approved by the Federal Drug Administration (FDA) to improve walking ability in people with PAD: pentoxifylline (1984) and cilostazol (1999). No new drugs have been approved since then to treat intermittent claudication. Furthermore, in recent studies in PAD patients, pentoxifylline does not significantly improve intermittent claudication symptoms or maximum walking distance over placebo. Recently published clinical practice guidelines recommend against prescribing pentoxifylline for intermittent claudication symptoms due to the lack of therapeutic benefit.

[0008]

[0009] Cilostazol improves treadmill walking capacity in people with symptomatic PAD Cilostazol is a phosphodiesterase inhibitor that improves walking ability by approximately 25% to 40%. Cilostazol is a phosphodiesterase type 3 inhibitor that acts by increasing the intracellular concentration of cyclic adenosine monophosphate, and in the process, the drug acts as a direct arterial vasodilator that inhibits platelet aggregation and improves blood perfusion. However, the mechanism by which cilostazol improves walking ability in PAD patients remains unclear.

[0009]

[0010] Side effects of cilostazol include headache, diarrhea, palpitations, and lightheadedness. There is a black box warning against prescribing cilostazol to subjects with a history of cardiovascular disease. Cilostazol should not be administered to PAD patients who also have heart failure. Cilostazol interacts with drugs that are regularly prescribed to patients with reduced renal function or cardiovascular disease, such as cinacalcet, clopidogrel, and ibandronate, thus increasing the risk of adverse reactions to these patients resulting from the combination of cilostazol with other drugs. Summary of the Invention [Problem to be solved by the invention]

[0010]

[0011] In conclusion, medical therapies for palliation of symptoms are limited, and although surgical or endovascular interventions are useful in some individuals, long-term results are often disappointing. As a result, there is a need to develop new, more effective and safer therapies to treat PAD. [Means for solving the problem]

[0011]

[0012] In a first aspect, the present invention provides a compound of general formula I, or a pharma- ceutically acceptable salt thereof, for use in increasing tissue perfusion and / or oxygenation in a subject in need thereof.

[0012] [ka]

[0013]

[0013] [wherein, R 1、 R 3、 R 5、 R 7、 R9 and R 11 are independently selected from OH, groups of formula II, III, IV:

[0014] [ka]

[0015] and heterologous moieties,

[0014] However: R, R, R, R, R and R 11 at least one of is selected from groups of formulas II, III and IV,

[0016] R1, R3, R5, R7, R9 and R 11 0, 1, 2 or 3 of the Regarding.

[0016]

[0017] In another aspect, the invention relates to a compound of general formula I as defined above for use in the treatment or prevention of ischemia in a subject in need thereof. In one version of this aspect, the invention relates to a compound of general formula I as described above for use in the treatment or prevention of an ischemia-related disease or condition in a subject in need thereof.

[0017]

[0018] In some embodiments, the present invention relates to compounds of general formula I as defined above, wherein the heterogeneous moiety is a group of formula V, a group of formula VI and a group of formula VII:

[0018] [ka]

[0019] is selected from n is an integer ranging from 2 to 200; R 13 is selected from H, methyl or ethyl.

[0020] In a further aspect, the present invention also relates to a compound of formula I, as defined above, which is The present embodiment also relates to a method for increasing tissue perfusion and / or oxygenation comprising administering to a subject in need thereof an amount of a compound of formula I, as defined above, together with a pharma- ceutical acceptable excipient or carrier. This embodiment may also be formulated as the use of a compound of formula I, as defined above, for the manufacture of a medicament for increasing tissue perfusion and / or oxygenation in a subject in need thereof.

[0021] In another aspect, the present invention also relates to a method for treating or preventing ischemia and / or ischemia-related diseases or conditions, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula I, as defined above, together with a pharma- ceutically acceptable excipient or carrier. This aspect may also be formulated as the use of a compound of formula I, as defined above, for the manufacture of a medicament for treating or preventing ischemia and / or ischemia-related diseases or conditions in a subject in need thereof.

[0022] In a further aspect, the present invention relates to a method for treating or preventing peripheral arterial disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula I, as defined above, together with a pharma- ceutically acceptable excipient or carrier. This aspect may also be formulated as the use of a compound of formula I, as defined above, for the manufacture of a medicament for treating or preventing peripheral arterial disease in a subject in need thereof.

[0023] The compounds of the present invention are particularly useful for increasing tissue perfusion and / or oxygenation in the lower limbs, in particular for the treatment or prevention of peripheral arterial disease (PAD) and closely related conditions, such as Critical Limb Ischemia (CLI).These compounds also exhibit many advantageous properties (e.g., a higher safety profile) compared to cilostazol, the currently indicated reference drug for the treatment of PAD.

[0024]

[0024] The present invention also provides a pharmaceutical composition comprising at least one compound of formula I, as defined above, for use in (i) increasing perfusion and / or oxygenation of tissue, (ii) treating or preventing ischemia and / or ischemia-related diseases, and / or (iii) treating or preventing PAD in a subject in need thereof. This aspect can also be formulated as the use of a pharmaceutical composition comprising at least one compound of formula I, as defined above, for the manufacture of a medicament for (i) increasing perfusion and / or oxygenation of tissue, (ii) treating or preventing ischemia and / or ischemia-related diseases, and / or (iii) treating or preventing PAD in a subject in need thereof. [Brief description of the drawings]

[0025] [Figure 1]

[0025] Figure 1 shows representative examples of inositol phosphate analogs in which two of the six X's are OPSO2 2- and the remaining X's are OSO3. Two specific forms of 4,6-di-(O-thiophosphate)-inositol-1,2,3,5-tetra-O-sulfate are shown. [Diagram 2]

[0026] FIG. 1 shows inositol phosphate analogs and derivatives that can be used to carry out the methods of the invention. The molecules shown are myo-inositol-pentaphosphate-2-PEG400, myo-inositol hexakissulfate, and scyllo-myo-inositol hexakissulfate. [Diagram 3]

[0027] 1 shows inositol phosphate analogs and derivatives that can be used to carry out the methods of the invention. X independently represents a phosphorus- and / or sulfur-containing group (e.g., phosphate, sulfate, or thiophosphate). R1 ​​represents a heterogeneous moiety (e.g., PEG or PG). [Figure 4]

[0028] FIG. 1 shows exemplary inositol phosphate analogs and derivatives that can be used to carry out the methods of the present invention. R1 represents a heterologous moiety (e.g., PEG or PG). n can be 2-200. [Diagram 5]

[0029] 1 shows exemplary inositol phosphate analogs and derivatives that can be used to practice the methods of the present invention. n can be from 2 to 200. [Figure 6-1]

[0030] 1 shows exemplary inositol phosphate analogs and derivatives that can be used to practice the methods of the present invention. n can be from 2 to 200. [Figure 6-2] Same as above [Figure 7]

[0031] FIG. 1 shows blood flow in the hind limbs of a rat model at D0 measured by Doppler laser imaging. Blood flow is shown in normalized perfusion units (PU). Normalization is obtained by comparing the raw data at D0 with the group 1 data. [Figure 8]

[0032] FIG. 1 shows blood flow in the hind limbs of a rat model at D6 measured by Doppler laser imaging. Blood flow is shown in normalized perfusion units (PU). Normalization is obtained by comparing the raw data at D6 with the Group 1 data. [Figure 9]

[0033] FIG. 1 shows blood flow in the hind limbs of a rat model at D12 measured by Doppler laser imaging. Blood flow is shown in normalized perfusion units (PU). Normalization is obtained by comparing the raw data at D12 with the Group 1 data. [Figure 10]

[0034] Figure 1 shows the percentage inhibition of aortic calcification in VitD rat model at D12. Calcium levels at the time of sacrifice were measured by ICP-OES. [Figure 11]

[0035] FIG. 1 shows blood flow in the hind limbs of a rat model at D12 and D18 (6 days after cessation of treatment) measured by Doppler laser imaging. Blood flow is shown in normalized perfusion units (PU). Normalization is obtained by comparing the raw data at D12 and D18 with the group 1 data. [Figure 12]

[0036] Figure 1 shows (A) Maximum walking distance (MWD) and (B) Maximum walking time (MWT) in a rat model at D10 measured by treadmill testing. Maximum walking distance is shown in meters (m) and maximum walking time is shown in minutes (min). [Figure 13]

[0037] Figure 1 shows the maximum walking distance (MWD) in a rat model at D17 (5 days after cessation of treatment) measured by treadmill testing. Maximum walking distance is shown in meters (m) up to a walking time of 40 min. [Figure 14]

[0038] Figure 1 shows the percentage inhibition of aortic calcification in the VitD rat model at D24 (12 days after cessation of treatment).Calcium levels at the time of sacrifice were measured by ICP-OES. [Figure 15]

[0039] FIG. 1 shows blood flow in the hind limbs of a rat model at D0, D5, and D13 (8 days after the start of treatment) measured by Doppler laser imaging. Blood flow is shown in normalized perfusion units (PU). Normalization is obtained by comparing the raw data at D0, D5, and D13 with the group 1 data. [Figure 16]

[0040] Figure 1 shows (A) Maximum walking distance (MWD) and (B) Maximum walking time (MWT) in a rat model at D11 (7 days after the start of treatment) measured by treadmill testing. Maximum walking distance is shown in meters (m) and maximum walking time is shown in minutes (min). [Figure 17]

[0041] Figure 1 shows the percentage inhibition of femoral artery calcification in VitD rat model at D13 (9 days after the start of treatment).Calcium levels at the time of sacrifice were measured by ICP-OES. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0026]

[0042] The present invention provides compounds, pharmaceutical compositions, methods and routes of administration for use in increasing tissue perfusion and / or oxygenation.The present invention also provides compounds, pharmaceutical compositions, methods and routes of administration for use in the treatment or prevention of ischemia and ischemia-related diseases and conditions.

[0027]

[0043] The compounds of the present invention are particularly useful for increasing tissue perfusion and / or oxygenation in the lower limbs, in particular for the treatment or prevention of peripheral arterial disease (PAD) and related conditions, such as critical limb ischemia (CLI).These compounds also exhibit many advantageous properties compared to other approved drugs for the treatment of PAD and CLI.

[0028] 1. Definitions of general terms and expressions

[0044] The invention includes embodiments in which only one member of the group is present in, employed in, or otherwise relevant to a given product or process. The invention also includes embodiments in which more than one, or all of the group members are present in, employed in, or otherwise relevant to a given product or process.

[0029]

[0045] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this description pertains.See, for example, Pei-Show J, Concise Dictionary of Biomedicine and Molecular Biology, 2nd Edition. (CRC Press, Boca Raton, FL, USA 2002); Lackie J, The Dictionary of Cell and Molecular Biology, 5th Edition. (Academic Press, Cambridge, MA, USA 2013) and Cammack R et al., Oxford Dictionary of Biochemistry and Molecular Biology, 2nd Edition, (Oxford University Press, Oxford, GB, 2006) provides the skilled artisan with a general dictionary that includes many of the terms used in this description.

[0030]

[0046] Units, prefixes, and symbols are based on their International System of Units (SI). Numerical ranges are inclusive of the numbers defining the range. When a range of values ​​is recited, it is to be understood that each intervening integer value, and each fraction thereof, between the recited upper and lower limits of that range is also specifically disclosed, along with each subrange between such values. The upper and lower limits of any range can be independently included or excluded from the range, and each range inclusive, exclusive, or inclusive of both limits is also encompassed within the scope of the invention.

[0031]

[0047] When a value is explicitly recited, it is to be understood that values ​​that are about the same content or amount as the recited value are also within the scope of the present invention. When a combination is disclosed, each subcombination of the elements of the combination is also specifically disclosed and is within the scope of the present invention. Conversely, when different elements or groups of elements are individually disclosed, their combinations are also disclosed. When any element of an invention is disclosed as having multiple alternatives of the invention, and each alternative is also disclosed herein, individually excluded or in any combination with other alternatives; two or more elements of an invention can have such an exclusion, and all combinations of the elements with such exclusion are disclosed herein.

[0032]

[0048] As used herein, the term "and / or" is to be construed as referring to each particular invention of the two specified features or components with or without the other. Thus, the term "and / or" used herein in the form of a phrase such as "A and / or B" is intended to include "A and B," "A or B," "A" (single), and "B" (single). Similarly, the term "and / or" used in the form of a phrase such as "A, B, and / or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (single); B (single); and C (single).

[0033]

[0049] As used herein and as applied to one or more values ​​of interest, the term "about" refers to a value similar to a specified reference value. In some embodiments, the term "about" refers to within a range of values ​​that is 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater or less than) the specified reference value, unless otherwise stated or otherwise clear from the context (except where such number exceeds 100% of possible values).

[0034]

[0050] As used herein, the term "critical limb ischemia" or "CLI" refers to severe blockage of arteries that significantly reduces blood flow to the limb and leads to severe pain, even skin ulcers, tenderness, or progression to gangrene.Critical limb ischemia is a very severe state of peripheral arterial disease.In some embodiments, administration of the inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention to a subject in need thereof improves the subject's ability to walk faster and longer distances compared to untreated cases.

[0035]

[0051] As used herein, the term "compound" is meant to include all isomers and isotopes of the depicted structures. As used herein, the term "isomer" means any geometric isomer, tautomer, zwitterion, stereoisomer, enantiomer, or diastereomer of a compound. A compound may contain one or more chiral centers and / or double bonds and therefore can exist as stereoisomers, e.g., double bond isomers (i.e., geometric E / Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers), etc. The present invention encompasses any and all isomers of the compounds described herein, including stereomerically pure forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) as well as enantiomeric and stereoisomeric mixtures, e.g., racemates. Enantiomeric and stereomeric mixtures of compounds and means for separating them into their component enantiomers or stereoisomers are well known. The compounds, salts, or complexes of the invention can be prepared in combination with solvents or water molecules to form solvates and hydrates by routine methods.

[0036]

[0052] As used herein, the term "cilostazol" refers to 6-[4-(1-cyclohexyl-1H-tetrazol-5yl)butoxy]-3,4-dihydro-2(1H)-quinolinone [CAS-73963-72-1], a quinolinone derivative that inhibits cellular phosphodiesterases. The molecular formula and weight of cilostazol are C, respectively. 20 H 27 N5O2 and 369.46 g / mol. Its structural formula is:

[0037] [ka]

[0038] It is.

[0053] As used herein, the term "effective amount" and the related terms "effective dosage" and "effective dose" with respect to (i) a compound of general formula I (e.g., inositol phosphate, inositol phosphate analogs, inositol phosphate derivatives, or combinations thereof), or (ii) a pharmaceutical composition comprising at least one of the compounds of item (i), is an amount sufficient to produce a beneficial or desired effect. In some embodiments, the beneficial or desired result is, for example, a clinical result, and thus, "effective amount" depends on the context in which it is applied. In the context of administering a therapeutic agent to enhance tissue perfusion and / or oxygenation, an effective amount of a therapeutic agent is, for example, an amount sufficient to (a) enhance tissue perfusion in a designated area, (b) halt, reduce, slow the progression of, or reverse ischemia in a designated area, or (c) improve mobility or walking ability (e.g., speed, distance) in a subject, when compared to the same parameters observed in the subject prior to administration of the therapeutic agent or in a population of control subjects not administered the therapeutic agent.

[0039]

[0054] As used herein, the term "ischemia" refers to a restriction in blood supply to a tissue resulting in a deficiency of oxygen required to maintain cellular metabolism. Ischemia includes not only insufficient oxygen, but also reduced availability of nutrients and inadequate removal of metabolic waste products. Ischemia can be partial (reduced perfusion) or complete.

[0040]

[0055] As used herein, the term "maximum walking distance" or "MWD" refers to the distance at which a subject becomes exhausted or in severe pain and is unable to continue walking unaided. In terms of assessing an increase in MWD, the increase is assessed by comparing the MWD values ​​of a subject before and after treatment with a therapeutic agent, or by comparing the MWD values ​​of the controls after treatment with a population of control subjects not treated with a therapeutic agent.

[0041]

[0056] As used herein, the term "maximal walking time" or "MWT" refers to the time at which a subject becomes exhausted or in severe pain and is unable to continue walking unaided. In terms of assessing an increase in MWT, the increase is assessed by comparing the MWT values ​​of a subject before and after treatment with a therapeutic agent, or by comparing the MWT values ​​of the controls after treatment with a population of control subjects not treated with a therapeutic agent.

[0042]

[0057] As used herein, the terms "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral and topical administration, usually by injection, and include, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injection and infusion (e.g., kidney dialysis infusion).

[0043]

[0058] As used herein, the term "peripheral arterial disease" or "PAD" refers to the narrowing of peripheral blood vessels to the legs (most commonly), stomach, arms, and head. Symptoms include intermittent claudication (e.g., leg pain when walking that resolves with rest), skin ulcers, bluish skin, cold skin, or poor nail and hair growth.

[0044]

[0059] As used herein, the terms "prevent," "preventing," and "prevention" refer to inhibiting the onset or reducing the occurrence of a disease or condition in a subject (e.g., avoiding the occurrence of ischemic tissue in a limb).

[0045]

[0060] As used herein, the term "SNF472" refers to an intravenous myo-inositol hexaphosphate hexasodium formulation. SNF472 is manufactured by dissolving myo-inositol hexaphosphate hexasodium in saline, followed by pH adjustment and sterile filtration. SNF472 is prepared in three different strengths: (i) 20 mg / mL and (ii) 90 mg / mL in 5 mL single-use vials (pH 5.8-6.2) formulated in saline, and (b) 30 mg / L in 10 mL single-use vials (pH 5.6-6.4) formulated in saline.

[0046]

[0061] As used herein, the term "subject", "individual", "animal" or "mammal" refers to any subject, particularly a mammalian subject, for which diagnosis, prognosis or therapy is desired. Mammalian subjects include, but are not limited to, humans, domestic livestock, farm livestock, zoo animals, sport animals, pet animals, such as dogs, cats, guinea pigs, rabbits, rats, mice, horses, cows, cows; primates, such as apes, monkeys, orangutans, and chimpanzees; canines, such as dogs and wolves; felines, such as cats, lions, and tigers; equines, such as horses, donkeys, and zebras; bears, food animals, such as cows, pigs, and sheep; ungulates, such as deer and giraffes; rodents, such as mice, rats, hamsters, and guinea pigs; and the like. In some embodiments, the subject is a human subject. In some embodiments, the subject is a human patient with reduced tissue perfusion and / or oxygenation in lower limb muscles or at risk of developing said condition. In some further embodiments, the subject is a human patient with ischemia and / or an ischemia-related disease or condition or at risk of developing said ischemia, ischemia-related disease or condition.

[0047]

[0062] As used herein, the term "substantially" refers to the qualitative condition of exhibiting a complete or nearly complete extent or degree of a desired characteristic or property. Those skilled in the art of biology understand that biological and chemical phenomena rarely, if ever, proceed to completion and / or to perfection or to reach or avoid absolute results. Thus, the term "substantially" is used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0048]

[0063] As used herein, the term "tissue perfusion" refers to the flow of blood or other perfusion fluid through the blood vessels of a particular tissue or organ. As used herein, "increased tissue perfusion" or "increasing tissue perfusion" refers to an increase in blood flow in a particular tissue region in a subject following administration of an inositol phosphate of the invention, compared to the same parameter observed in the subject prior to administration of said therapeutic agent or in a population of control subjects not administered said therapeutic agent.

[0049]

[0064] As used herein, the term "treat" or "treatment" refers to the administration of a compound or pharmaceutical composition of the present invention to (i) slow, (ii) inhibit, (iii) halt, or (iv) reverse the progression of a disease or condition after clinical signs appear. Control of disease progression includes, but is not limited to, However, it is understood that the term "treatment" refers to beneficial or desired clinical results, including alleviation of symptoms, shortening of disease duration, stabilization of pathological condition (particularly to avoid further deterioration), delay of disease progression, improvement of pathological condition, and remission (partial and complete). Control of disease progression also includes prolongation of survival time compared to the predicted survival time if no treatment was applied. In the context of the present invention, the term "treat" or "treatment" specifically refers to (a) increasing tissue perfusion and / or oxygenation, or (b) stopping, reducing, slowing or reversing the development of ischemic tissue, particularly in the lower limbs, or (c) improving mobility or walking ability (e.g., speed, distance, endurance) in subjects administered with the compound or pharmaceutical composition of the present invention.

[0050]

[0065] As used herein, the term "ambulatory ability" refers to the ability of a subject to move autonomously without assistance. The parameters MWD and MWT indicate the ambulatory ability of a subject.

[0051] 2.Compound

[0066] The compounds for use in the present invention are inositol phosphates, as defined in the first aspect of the present invention, and their analogs and derivatives. As used herein, the term "inositol phosphate" refers to a compound having an inositol ring and 1, 2, 3, 4, 5 or 6 phosphate groups, or a combination thereof. Myo-inositol hexaphosphate (IP6) is an exemplary inositol phosphate of the present invention. In some embodiments, the inositol phosphate is pure (e.g., more than 99% of the inositol phosphate species are the same species, e.g., IP6) or substantially pure (e.g., at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the inositol phosphate species are the same species, e.g., IP6). In some embodiments, the inositol phosphate is a mixture, e.g., including variable amounts of IP1, IP2, IP3, IP4, IP5, and IP6. In some embodiments, the inositol phosphates are a racemic mixture.

[0052]

[0067] The present invention also contemplates inositol phosphate analogs.As used herein, "inositol phosphate analogs" refers to compounds that have rings with different numbers of carbons on the inositol ring (i.e., 5 or 7 carbons) and / or have at least one sulfate or thiophosphate group.For example, compounds that include rings with 5, 6, or 7 carbons and at least one phosphate, sulfate, or thiophosphate group should be considered to be inositol phosphate analogs.

[0053]

[0068] As used herein, the term "inositol phosphate derivative" refers to an inositol phosphate or inositol phosphate analogue that contains a heterogeneous moiety (i.e., a group that is not a phosphate, sulfate, or thiophosphate). For example, inositol pentasulfate containing a polyethylene glycol heterogeneous moiety, or myo-inositol hexaphosphate containing a polyglycerin heterogeneous moiety, would be considered an inositol phosphate derivative.

[0054]

[0069] As used herein, the term "heterogeneous moiety" refers to a group in a compound of formula I that is not a phosphate, sulfate, or thiophosphate and that confers desirable properties to such a compound. For example, a heterogeneous moiety (e.g., polyglycerin or polyethylene glycol) can increase the solubility of a compound. In some embodiments, a heterogeneous moiety can confer multiple desirable properties (e.g., polyglycerin and polyethylene glycol can increase the solubility of a compound and decrease the clearance rate of a compound).

[0055]

[0070] As used herein, the terms "inositol phosphate of the present invention" and "inositol phosphate of the present invention" are generic terms that include "inositol phosphate", "inositol phosphate analogs", "inositol phosphate derivatives" and combinations thereof. In some embodiments, the term "inositol phosphate of the present invention" includes compositions that include "inositol phosphate", "inositol phosphate analogs", and "inositol phosphate derivatives" or combinations thereof, and at least one additional therapeutic agent. In some embodiments, the additional therapeutic agent includes cilostazol, pentoxifylline, or a combination thereof.

[0056]

[0071] Compounds of the invention that contain a ring with 5, 6, or 7 carbons and at least one sulfate or thiophosphate group, but no phosphate group, would be considered to be "inositol phosphate analogs" or "inositol phosphate analogs" in the context of the invention. Thus, the term "inositol phosphates of the invention" encompasses not only phosphate-containing compounds, but also compounds that contain a ring with 5, 6, or 7 carbons and at least one sulfate or thiophosphate group, but no phosphate group.

[0057]

[0072] Representative inositol phosphates of the present invention are shown in Figures 1-6. Figure 3 shows numerous examples of inositol phosphates, all of which are in the form of the myo conformation. In addition to myo-inositol, other naturally occurring stereoisomers of inositol are scyllo-, muco-, 1D-chiro-, 1L-chiro-, neo-inositol, allo-, epi-, and cis-inositol. As their names suggest, 1L- and 1D-chiro-inositol are the only pair of inositol enantiomers, but they are enantiomers of each other and not of myo-inositol. It is understood that any exemplary inositol phosphates shown in the present disclosure are not limited to the representative conformations shown. Thus, for example, the example shown in Figure 3 should also encompass the corresponding equivalents in the scyllo-, muco-, 1D-chiro-, 1L-chiro-, neo-inositol, allo-, epi-, and cis-inositol conformations. In its most stable conformation, the myo-inositol isomer adopts a chair conformation that moves the greatest number of hydroxyls to equatorial positions, which are the furthest apart from each other. In this conformation, the naturally occurring myo-isomer has a structure in which five of the six hydroxyls (the first, third, fourth, fifth, and sixth) are equatorial, while the second hydroxyl group is axial.

[0058] [ka]

[0059] 2.1. Inositol Phosphates, Analogues and Derivatives

[0073] In some embodiments, R of the compound of formula I 1、 R 3、 R 5、 R 7、 R9 and R 11 at least one of independently represents H, -X, -OX, -NHX, -NX2, -SX, -OSO3HX, -OSO3X2, or a compound of formula II, formula III, or formula IV, and each X is independently H, C 1~30 Alkyl, C 2~30 alkynyl or Cy1, C 1~30 Alkyl, C2~30 Alkenyl and C 2~30 Alkynyl is independently one or more R 14 and Cy1 is optionally substituted with one or more R 15 Cy1 represents a 3- to 10-membered carbocyclic or heterocyclic ring, which may be saturated, partially unsaturated or aromatic, said heterocyclic ring having 1 to 4 heteroatoms selected from among O, S and N, said ring may be bonded to the remainder of the molecule via any available C atom, Cy1 is optionally fused to 1 to 4 5- or 6-membered rings, each of which may be saturated, partially unsaturated or aromatic, carbocyclic or heterocyclic, said fused heterocyclic rings may contain 1 or 2 heteroatoms selected from among O, N and S; each R 13 are independently H, C 1~30 Alkyl, -NH2, -NHC 1~30 Alkyl or N(C 1~30 alkyl)2, each C 1~30 The alkyl is optionally substituted, independently, with one or more halogen, -OH, -CN, and -NO groups; each R 14 and R 15 are independently -OH, C 1~30 Alkoxy, C 1~30 Alkylthionyl, C 1~30 Acyloxy, phosphate, halogen, trihalo C 1~30 Represents alkyl, nitrile azide.

[0060]

[0074] In some implied embodiments, each X is independently H, C 1~30 represents alkyl or Cy1, C 1~30 Alkyl is one or more R 14 Cy1 is optionally substituted by one or more R 15 each R 14 and R 15 are independently -OH, C 1~30 Alkoxy, C 1~30 Alkylthionyl, C 1~30Acyloxy, phosphate, halogen, trihalo C 1-30 Alkyl, Ni In some embodiments, each X represents H, C, 1~30 In some embodiments, each X represents H.

[0061]

[0075] In some additional embodiments, R1, R3, R5, R7, R9 and R 11 At least one of the groups independently represents a compound of formula II, formula III or formula IV, and each R 13 are independently H, C 1~30 Alkyl, -NH2, -NHC 1~30 Alkyl or -N(C 1~30 alkyl)2, each C 1~30 The alkyl is optionally substituted independently with one or more halogen, -OH, -CN, and -NO2 groups; R2, R4, R6, R8, R 10 and R 12 each independently represents H.

[0062]

[0076] In a further embodiment, R1, R3, R5, R7, R9 and R 11 independently represents a compound of formula II, formula III, or formula IV, and each R 13 are independently H or C 1~30 Represents alkyl, and each C 1~30 The alkyl is optionally substituted independently with one or more halogen, -OH, -CN, and -NO2 groups; R2, R4, R6, R8, R 10 and R 12 each independently represents H.

[0063]

[0077] In a further embodiment, R1, R3, R5, R7, R9 and R 11 at least one of R represents a compound of formula II, formula III, or formula IV; 13 are independently H or C 1~30 In another embodiment, R1, R3, R5, R7, R9 and R 11at least one of R represents a compound of formula II, formula III or formula IV; 13 represents H.

[0064]

[0078] In particular embodiments, the compound is inositol hexaphosphate (IP6). In other embodiments, the compound is inositol monophosphate (IP1), inositol diphosphate (IP2), inositol triphosphate (IP3), inositol tetraphosphate (IP4), or inositol pentaphosphate (IP5). In some embodiments, the compound includes combinations of IP1, IP2, IP3, IP4, IP6, and IP6. In some embodiments, IP6 can form other inositol phosphates (IP5, IP4, IP3, IP2, IP1) by dephosphorylation in vivo. Inositol is taken to mean any isomeric form of the molecule, for example, myo-inositol.

[0065]

[0079] In some embodiments, the compounds for use in the present invention have formula I,

[0080] R7 is OSO3 - and R9, R5, R3, R1 and R 11 is OPO3 2- , OPSO2 2- or OSO3 - are independently selected from;

[0081] R9, R5 and R1 are OPO3 2- and R7, R3 and R 11 is OSO3 - and;

[0082] R9, R5 and R1 are OSO3 - and R7, R3 and R 11 is OPO3 2- and;

[0083] R3, R1 and R 11 is OSO3 - R9, R7 and R5 are OPO3 2- and;

[0084] R3, R1 and R11 is OPO3 2- R9, R7 and R5 are OSO3 - and;

[0085] R7 and R1 are OPO3 2- and R9, R5, R3, and R 11 is OPO3 2- and;

[0086] R7 and R1 are OSO3 - and R9, R5, R3, and R 11 is OPO3 2- and;

[0087] R7 and R5 are OPO3 2- and R9, R3, R1, and R 11 is OSO3 - or

[0088] R7 and R5 are OSO3 - and R9, R3, R1, and R 11 O PO3 2- [is].

[0066]

[0089] The inositol phosphates of the present invention also include compounds produced as metabolites during physiological dephosphorylation (or, in the case of compounds containing sulfate or thiophosphate groups, desulfation or dethiosulfation).

[0067]

[0090] In some embodiments, the compounds administered in doses according to the methods disclosed herein are prodrugs that, after hydrolysis or other intracellular or extracellular processing, produce the inositol phosphates of the invention.

[0068]

[0091] The inositol phosphates of the present invention also include any combination of the inositol phosphates, inositol phosphate analogs, and derivatives thereof disclosed herein.

[0092] All compounds of formula I contain a group with a COP or COS bond, which gives the compound an affinity to calcium-containing crystals and a bond that is sufficiently unstable to be hydrolyzed in vivo, thereby preventing irreversible binding to calcium-containing crystals, such as hydroxyapatite (HAP) in bone. If such irreversible binding were to occur, it would have a negative effect on bone remodeling, as do bisphosphonates when administered long-term, because they contain a PCP bond that cannot be hydrolyzed by the body. At the other extreme, phosphorylated compounds that do not contain the COP bond, such as pyrophosphate, the POP bond means that they are too easily hydrolyzed in the intestinal tract, and therefore only parenteral administration is feasible. Compounds of the present invention with COP bonds, COS bonds, and combinations thereof, represent a good middle ground due to their effectiveness and the fact that the body shows mechanisms for excreting the compounds and therefore reducing the risk of side effects (e.g., compounds with PCP bonds can show a half-life of several months in vivo, thereby affecting, for example, bone remodeling).

[0069]

[0093] The term "alkyl" or "alkyl group" in the context of the present invention means a saturated hydrocarbon moiety, which may be linear, branched, cyclic or cyclic with linear or branched side chains. The term alkyl includes partially unsaturated hydrocarbons, such as propenyl. Examples are methyl, ethyl, n- or isobutyl, n- or cyclohexyl. The term alkyl can be extended to alkyl groups linked or bridged by heteroatoms. Heteroatoms in the context of the present invention are nitrogen (N), sulfur (S) and oxygen (O).

[0070]

[0094] An "amine functional group" or "amine group" is a NR'R" functional group, where R' and R" are independently selected from hydrogen and C1-C5 alkyl. In some embodiments, R' and R" are selected from hydrogen and C1-C3 alkyl. A "hydroxy functional group" or "hydroxy group" is OH.

[0071]

[0095] A "thiol functional group" or "thiol group" is SH. A "carboxylic acid functional group" or "carboxylic acid group" is COOH or its anion, COO - A "carboxylic acid amide" is CONR'R'', where R' and R'' have independently the meanings given above. A "sulfonic acid" is SO3H. A "sulfonic acid amide" is SO2NR'R'', where R' and R'' have independently the meanings given above.

[0072]

[0096] "C1-C3 alkyl" in the context of the present invention means a saturated linear or branched hydrocarbon having 1, 2, or 3 carbon atoms, in which one carbon-carbon bond may be unsaturated and one CH2 moiety may be replaced with oxygen (ether bridge). Non-limiting examples of C1-C3 alkyl are methyl, ethyl, propyl, propane, phenyl ... -enyl and prop-2-ynyl.

[0073]

[0097] "C1-C5 alkyl" in the context of the present invention means a saturated linear or branched hydrocarbon having 1, 2, 3, 4 or 5 carbon atoms, in which one or two carbon-carbon bonds may be unsaturated and one CH2 moiety may be replaced with oxygen (ether bridge). Non-limiting examples for C1-C5 alkyl include the examples given above for C1-C3 alkyl, as well as n-butyl, 2-methylpropyl, tert-butyl, 3-methylbut-2-enyl, 2-methylbut-3-enyl, 3-methylbut-3-enyl, n-pentyl, 2-methylbutyl, 3-methylbutyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 1,2-dimethylpropyl, but-3-enyl, but-3-ynyl and pent-4-ynyl. "C3-C 10 "Alkyl" means a saturated straight or branched chain hydrocarbon having 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, in which 1, 2, or 3 carbon-carbon bonds may be unsaturated and one CH2 moiety may be replaced with oxygen (ether bridge).

[0074]

[0098] The term "C" as a group or part of a group 1~30 "Alkyl" means straight or branched chain alkyl groups containing from 1 to 30 carbon atoms, including methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, decyl, and dodecyl groups, among others.

[0075]

[0099] The term “C 2~30 "Alkenyl" means a straight or branched alkyl chain containing 2 to 30 carbon atoms and also containing one or more double bonds. Examples include ethenyl, 1-propenyl, 2-propenyl, isopropenyl 1-butenyl, 2-butenyl, 3-butenyl, and 1,3-butadienyl, among others.

[0076]

[0100] The term “C 2~30"Alkynyl" means a straight or branched alkyl chain containing 2 to 30 carbon atoms and also containing one or more triple bonds. Examples include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, and 1,3-butadiynyl, among others.

[0077]

[0101] "Cy1 group" means a 3- to 10-membered carbocyclic or heterocyclic ring, which may be saturated, partially unsaturated or aromatic, and is bonded to the remainder of the molecule by any available C atom. When it is a heterocyclic ring, Cy1 contains 1 to 4 heteroatoms selected from among N, O, and S. Furthermore, Cy1 may be optionally fused with a 4, 5, or up to 6-membered carbocyclic or heterocyclic ring, which may be saturated, partially unsaturated or aromatic. When the fused ring is a heterocyclic ring, said ring contains 1 or 2 heteroatoms selected from among N, O, and S. Examples of Cy1 include, among others, phenyl, naphthyl, thienyl, furyl, pyrrolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, 1,3,4-thiadiazolyl, 1,2,4-thiadiazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, benzimidazolyl, benzofuranyl, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, benzothiazolyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, azetidinyl and aziridinyl.

[0078]

[0102] "C" as a group or part of a group 1~30 "Alkoxy group" means -OC 1~30 means an alkyl group, C 1~30 The alkyl moiety has the same meaning as above. Examples include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy and tert-butoxy.

[0079]

[0103] "C" as a group or part of a group 1~30"Alkylthionyl group" means -SOC 1~30 means an alkyl group, C 1~30 The alkyl portion has the same meaning as above. Examples include methylthionyl, ethylthionyl, propylthionyl, isopropylthionyl, butylthionyl, isobutylthionyl, sec-butylthionyl and tert-butylthionyl.

[0080]

[0104] "C" as a group or part of a group 1~30 "Acyloxy group" means -COC 1~30 means an alkyl group, C 1~30 The alkyl moiety has the same meaning as above. Examples include acetyl, ethanoyl, propanoyl and 2,2-diisopropylpentanoyl.

[0081]

[0105] The term "halogen" or its abbreviation halo means fluorine, chlorine, bromine and iodine.

[0106] "Trihalo C 1~30 "Alkyl group" means a C alkyl group, as defined above, with three halogen groups. 1~30 means the group resulting from the replacement of three hydrogen atoms of an alkyl group. Examples include trifluoromethyl, tribromomethyl, trichloromethyl, triiodomethyl, trifluoroethyl, tribromoethyl, trichloroethyl, triiodoethyl, tribromopropyl, trichloropropyl, and triiodopropyl, among others.

[0082]

[0107] -NHC 1~30 "Alkyl group" is defined as above. 1~30 It means the group resulting from the replacement of one hydrogen atom of a -NH2 group by an alkyl group. Examples include methylamine, ethylamine, propylamine, butylamine and pentylamine, among others.

[0083]

[0108] "-N(C 1~30 "Alkyl group" is defined as above. 1~30It means the group resulting from the replacement of the two hydrogen atoms of a -NH group by an alkyl group. Examples include dimethylamine, diethylamine, diisopropylamine, dibutylamine and diisobutylamine, among others.

[0084]

[0109] The phrase "optionally substituted with one or more" refers to the possibility that a group can be substituted with one or more (e.g., 1, 2, 3, or 4) substituents. In some embodiments, if the group has enough available positions that can be substituted, the group can be substituted with 1, 2, or 3 substituents, and even 1 or 2 substituents. If present, the substituents can be the same or different and can be at any available position.

[0085]

[0110] In some embodiments, the inositol phosphates of the invention include those compounds disclosed in WO2017098033 and WO2017098047, and US US9358243. In some embodiments, the inositol phosphates of the invention used include those compounds disclosed in Figures 1-6.

[0086]

[0111] In some embodiments, the inositol phosphates, inositol phosphate analogs, and derivatives thereof are represented by formula (VIII), formula (IX), or formula (X):

[0087] [ka]

[0088] This includes compounds of the formula:

[0112] [Wherein each X is independently OPO3 2- , OPSO2 2- , or OSO3 - Z is an alkyl chain containing 1 to 3 carbon and / or heteroatoms and optionally containing an X group, X also being OPO3 2- , OPSO2 2- , or OSO3 - Also selected from;R1 is an optional heterologous moiety (see Section 2.2. below). In some embodiments, a molecule comprises two or more heterologous moieties, in which case the heterologous moieties may be the same or different.

[0089]

[0113] In some embodiments, Z, as used in formula (VIII), is CH, CHX, CHR 1 , CXR 1 , CH2-CH2, CH2-CHX, CHX-CHX, CHR 1 -CHX, CXR 1 -CHX, CHR 1 -CH2, CXR 1 -CH2, CHR 1 -CHOH, CH2-CH2-CH2, CH2-O-CH2, CHOH-CH2-CH2, CHOH-CHOH-CHR 1 , CHOH-CHR 1 -CHOH, CHX-CH2-CH2, CH2-CHX-CH2, CHX-CHX-CH2, CHX-CH2-CHX or CHX-CHR 1 -CHX, where X is independently OPO3 2- , OPSO2 2- , and OSO3 - is selected from.

[0090]

[0114] In some embodiments, Z, as used in formula (VIII), is (CHX) p CHX(CHX) q p and q each independently have a value from 0 to 2, with the proviso that (p+q) has a value of 0, 1, or 2; one, two, or three Xs may be heterogeneous moieties (e.g., PEG), and the remaining Xs are OPO3 2- , OPSO2 2- , and OSO3 - In some embodiments, all X in Z are OPO3. 2- In some embodiments, all X of Z are OSO3. - This is not necessarily the case.

[0091]

[0115] In some embodiments, one, two, or three of the X's in the compound of formula (VIII), formula (IX), or formula (X) can be heterogeneous moieties, with the remaining X's being OPO3 2- , OPSO2 2- , or OSO3 - can be independently selected from

[0092]

[0116] The above formula (VII) describes a 5-, 6-, or 7-membered alkyl ring, with one or more optional heterostructural moieties attached to one of the carbon atoms forming the ring.

[0093]

[0117] In some embodiments, for example, the inositol phosphates, inositol phosphate analogs, and derivatives thereof used in the methods and compositions disclosed herein include compounds of formula (XI) or formula (XII).

[0094] [ka]

[0095]

[0118] [In the formula,

[0119] X 2 is OSO3 - and X 1 , X 3 , X 4 , X 5 and X 6 is OPO3 2- , OPSO2 2- or OSO3 - are independently selected from;

[0120] X 1 , X 3 and X 5 is OPO3 2- and X 2 , X 4 and X 6 is OSO3 - and;

[0121] X 1 , X 3 and X5 is OSO3 - and X 2 , X 4 and X 6 is OPO3 2- and;

[0122] X 4 , X 5 and X 6 is OSO3 - and X 1 , X 2 and X 3 is OPO3 2- and;

[0123] X 4 , X 5 and X 6 is OPO3 2- and X 1 , X 2 and X 3 is OSO3 - and;

[0124] X 2 and X 5 is OPO3 2- and X 1 , X 3 , X 4 , and X 6 is OPO3 2- and;

[0125] X 2 and X 5 is OSO3 - and X 1 , X 3 , X 4 , and X 6 is OPO3 2- and;

[0126] X 2 and X 3 is OPO3 2- and X 1 , X 4 , X 5 , and X 6 is OSO3 - or

[0127] X 2 and X 3 is OSO3- and X 1 , X 4 , X 5 , and X 6 is OPO3 2- [is].

[0096]

[0128] In some embodiments, the inositol phosphates or their metabolites of the present invention can be detected and / or quantified using the methods disclosed in US 9612250. See also US 8377909, US 8778912, and US 20070066574.

[0097]

[0129] The compounds disclosed herein can be in any form commonly used in pharmaceutical technology.Detailed embodiments include, but are not limited to, sodium salt, magnesium salt, potassium salt, ammonium salt, free acid, or mixtures of the above.Other pharmaceutically acceptable salts are known to those skilled in the art and can be easily obtained.In detailed embodiments, the compound for use as defined in the first embodiment of the present invention is sodium salt, for example, hexasodium inositol hexaphosphate.

[0098]

[0130] The present invention also contemplates sodium salts of inositol monophosphate, inositol diphosphate, inositol triphosphate, inositol tetraphosphate and inositol pentaphosphate in any of the forms of inositol isomers, particularly in myo-inositol.A specific example of a compound for use in the present invention is myo-inositol hexaphosphate hexasodium salt.Sodium salts offer several advantages in terms of the production of the resulting IP6 formulation and the level of impurities in the resulting IP6 formulation.

[0099] 2.2. Heterogeneous Structure

[0131] In some embodiments, the present invention relates to compounds of general formula I as defined above, wherein the heterogeneous moiety is a group of formula V, a group of formula VI and a group of formula VII:

[0100] [ka]

[0101] is selected from

[0132] n is an integer ranging from 2 to 200; R 13 is selected from H, methyl or ethyl.

[0102]

[0133] In some embodiments, the compounds for use in the present invention, such as the inositol phosphate derivatives of the present invention, can include one or more groups selected from the groups of formula V, VI and VII. These groups are heterogeneous structural moieties that confer advantageous properties with respect to the corresponding molecules lacking such one or more heterogeneous structural moieties. Examples of such advantageous properties that can be conferred to inositol phosphate or inositol phosphate analogs by heterogeneous structural moieties or combinations thereof include, but are not limited to, (a) increased solubility, (b) reduced degradation or metabolic rate, (c) increased plasma half-life, (d) reduced hepatic metabolic rate, (e) reduced clearance rate, (f) reduced toxicity, (g) reduced irritability and (h) reduced side effects, among others. These advantageous properties can be evaluated or quantified using methods known in the art without undue experimentation.

[0103]

[0134] In some embodiments, the heterogeneous moiety is, for example, polyethylene glycol (PEG) or polyglycerol (PG). Thus, in some embodiments, the compound for use in the present invention is any of the compounds defined in the above disclosed embodiments that include a heterogeneous moiety, i.e., one of the groups of formula I is selected from the groups of formula V, VI and VII. In some embodiments, the heterogeneous moiety includes polyethylene glycol (PEG). In some embodiments, the heterogeneous moiety consists of polyethylene glycol, i.e., R1, R3, R5, R7, R9 and R of the compound of formula I according to the first embodiment of the present invention are selected from the groups of formula V, VI and VII. 11At least one of R1, R3, R5, R7, R9 and R9 of the compound of formula I according to the first aspect of the present invention is a group of formula V. Alternatively, the heterogeneous moiety comprises polyglycerol. In some embodiments, the heterogeneous moiety is composed of polyglycerol, i.e., R1, R3, R5, R7, R9 and R9 of the compound of formula I according to the first aspect of the present invention are 11 At least one of the groups is selected from the group of formula VI or VII. In another embodiment, the compound of formula I according to the first aspect of the invention is or VII groups, e.g., two PEGs (groups of formula V), three PEGs, three polyglycerols (groups of formula VI), three PGs, or any combination thereof, e.g., one PEG and one PG, or two PEGs and one polyglycerol. In some embodiments, all of the residues of formula I (i.e., those that are not groups selected from V, VI, and VII) are groups selected from II, III, and IV. In some embodiments, the compound of formula I according to the first aspect of the invention contains two groups selected from groups of formula VI or VII, e.g., two PEGs (groups of formula V) or three polyglycerols (groups of formula VI) or one PEG and one polyglycerol, and the residues are all groups of formula II. In some embodiments, R3 and R7 of the compound of formula I are selected from groups of formula V, VI, and VII. In some embodiments, R3 and R7 of the compound of formula I are groups of formula V, and R1, R5, R9 and R of the compound of formula I are groups of formula V 11 is a group of formula II.

[0104]

[0135] The groups of formulae V, VI and VII are R 13 ═H, methyl or ethyl, and n is an integer from 2 to 200. 13In particular embodiments, n is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59 ,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 1 58, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 189, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, or 200.

[0105]

[0136] In some embodiments, n is 2 to 10, 10 to 20, 20 to 30, 30 to 40, 40 to 50, 50 to 60, 60 to 70, 70 to 80, 80 to 90, 90 to 100, 100 to 110, 110 to 120, 120 to 130, 130 to 140, 140 to 150, 150 to 160, 160 to 170, 170 to 180, 180 to 190, or 190 to 200.

[0106]

[0137] In some specific embodiments, n has a value from 2 to 200, from 2 to 20, from 10 to 30, or from 9 to 45.

[0138] In some embodiments, the PEG is a branched PEG having 3 to 10 PEG chains emanating from a central core group.

[0107]

[0139] In some embodiments, the PEG moiety is a monodisperse polyethylene glycol. In the context of the present invention, a monodisperse polyethylene glycol (mdPEG) is a PEG having a single, defined chain length and molecular weight. mdPEG is usually produced by separation from the polymerization mixture by chromatography. In some formulas, the monodisperse PEG moiety is given the abbreviation mdPEG. In some embodiments, the PEG is a Star PEG. Star PEG has 10 to 100 PEG chains extending from a central core group.

[0108]

[0140] In some embodiments, the PEG is a Comb PEG, which has multiple PEG chains typically grafted onto a polymer backbone.

[0141] In some embodiments, the PEG has a molar mass of 100 g / mol to 3000 g / mol, in particular 100 g / mol to 2500 g / mol, and more particularly about 100 g / mol to 2000 g / mol. In some embodiments, the PEG has a molar mass of 200 g / mol to 3000 g / mol, in particular 300 g / mol to 2500 g / mol, and more particularly about 400 g / mol to 2000 g / mol.

[0109]

[0142] In some embodiments, the PEG is 100 , PEG 200 , PEG 300 , PEG 400 , PEG 500 , PEG 600 , PEG 700 , PEG 800 , PEG 900 , PEG 1000 , PEG 1100, PEG 1200 , PEG 1300 , PEG 1400 , PEG 1500 , PEG 1600 , PEG 1700 , PEG 1800 , PEG 1900 , PEG 2000 , PEG 2100 , PEG 2200 , PEG 2300 , PEG 2400 , PEG 2500 , PEG 1600 , PEG 1700 , PEG 1800 , PEG 1900 , PEG 2000 , PEG 2100 , PEG 2200 , PEG 2300 , PEG 2400 , PEG 2500 , PEG 2600 , PEG 2700 , PEG 2800 , PEG 2900 , or PEG 3000 In one particular embodiment, the PEG is 400 In another detailed embodiment, the PEG is 2000 It is.

[0110]

[0143] In other particular embodiments, R3 and / or R7 of the compound of formula I are groups of formula V, R 13 is H and n is an integer from 9 to 45. In other detailed embodiments, R3 and R7 of the compound of formula I are groups of formula V, R 13 is H, n is an integer from 9 to 45, and R1, R5, R9 and R 11 are all groups of formula II. In other particular embodiments, the compound of formula I has R3 and R7 = groups of formula V, 13 is H, n is an integer from 9 to 45, and R1, R5, R9 and R 11 are all groups of formula II.

[0111]

[0144] In some other embodiments, the heterogeneous moiety has the formula ((R-O-(CH-CHOH-CHO) n -), where R3 is hydrogen, methyl or ethyl, and n has a value from 3 to 200. In some embodiments, n has a value from 3 to 20. In some embodiments, n has a value from 10 to 30. In some alternatives of these embodiments, n has a value from 9 to 45. In some embodiments, the heterogeneous moiety is a polyglycerol (PG) described by the formula (R 3 -O-(CH2-CHOR 5 -CH2-O) n -) [where R 5 or a branched polyglycerol described by the formula (R 3 -O-(CH2-CHOH-CH2-O) n -) [where R 3 is hydrogen, methyl or ethyl. In some embodiments, the heterologous moiety is represented by the formula (R 3 -O-(CH2-CHOR 5 -CH2-O) n -) [where R 5 is hydrogen] or a hyperbranched polyglycerol described by the formula (R 3 -O-(CH2-CHOR 6 -CH2-O) n -) [where R 6 is hydrogen] or a glycerol chain described by the formula (R 3 -O-(CH2-CHOR 7 -CH2-O) n -) [where R 7 is hydrogen] or a glycerol chain described by the formula (R 3 -O-(CH2-CHOH-CH2-O) n -) [where R 3is hydrogen, methyl or ethyl. Hyperbranched glycerols and methods for their synthesis are known in the art. See Oudshorn M et al., Biomaterials 2006;27:5471-5479, Wilms D et al., Acc Chem Res 2010;43:129-141 and references cited therein.

[0112]

[0145] In some embodiments, PG has a molar mass between 100 g / mol and 3000 g / m ol, in particular 100 g / mol to 2500 g / mol, more particularly approximately 100 g / mol to 2000 g / mol. In some embodiments, PG has a molar mass of 200 g / mol to 3000 g / mol, in particular 300 g / mol to 2500 g / mol, more particularly approximately 400 g / mol to 2000 g / mol.

[0113]

[0146] In some embodiments, PG is 100 , P.G. 200 , P.G. 300 , P.G. 400 , P.G. 500 , P.G. 600 , P.G. 700 , P.G. 800 , P.G. 900 , P.G. 1000 , P.G. 1100 , P.G. 1200 , P.G. 1300 , P.G. 1400 , P.G. 1500 , P.G. 1600 , P.G. 1700 , P.G. 1800 , P.G. 1900 , P.G. 2000 , P.G. 2100 , P.G. 2200 , P.G. 2300 , P.G. 2400 , P.G. 2500 , P.G. 1600 , P.G. 1700 , P.G. 1800 , P.G. 1900 , P.G. 2000 , P.G. 2100 , P.G. 2200 , P.G.2300 , P.G. 2400 , P.G. 2500 , P.G. 2600 , P.G. 2700 , P.G. 2800 , P.G. 2900 , or P.G. 3000 In a particular embodiment, the PG is 400 In another detailed embodiment, the PG is 2000 It is.

[0114]

[0147] In other particular embodiments, R3 and / or R7 of the compound of formula I are groups of formula VI, R 13 is H and n is an integer from 9 to 45. In other detailed embodiments, R3 and R7 of the compound of formula I are groups of formula VI, R 13 is H, n is an integer from 9 to 45, and R1, R5, R9 and R 11 are all groups of formula II. In other particular embodiments, the compounds of formula I have R3 and R7 = groups of formula VI, 13 is H, n is an integer from 9 to 45, and R1, R5, R9 and R 11 are all groups of formula II.

[0115] 3. Pharmaceutical Compositions

[0148] In another aspect, the present invention also refers to pharmaceutical compositions comprising the compound defined in any of the above disclosed aspects. In some aspects, the pharmaceutical compositions comprise the compound defined in any of the above disclosed aspects together with one or more pharma- ceutically acceptable excipients or carriers. These pharmaceutical compositions are for use in increasing tissue perfusion and / or oxygenation in subjects who require such an increase. In some aspects, these pharmaceutical compositions are for use in treating or preventing ischemia and / or ischemia-related diseases or conditions. In some aspects, the pharmaceutical compositions of the present invention are used for treating or preventing PAD or CLI.

[0116]

[0149] As used herein, the term "excipient" refers to a substance that aids in the absorption of, stabilizes, activates, or aids in the preparation of a pharmaceutical composition. Thus, examples of excipients used in parenteral formulations include, but are not limited to, antibacterial agents (e.g., benzalkonium chloride, metacresol, thimerosal), co-solvents (e.g., ethanol), buffers, and pH regulators (e.g., carbonate, citrate, phosphate solutions).

[0117]

[0150] As in the case of excipients, a "pharmaceutically acceptable vehicle" is a substance used in a composition to dilute any of the components contained therein to a determined volume or weight. A pharmaceutically acceptable vehicle is an inactive substance or a substance that has a similar action to any of the elements that comprise the pharmaceutical composition of the present invention. The role of the vehicle is to allow the incorporation of other elements, to allow better dosing and administration, or to provide consistency and shape to the composition.

[0118]

[0151] The pharmaceutical composition can comprise from about 1% to about 95% of the compound defined in any of the above disclosed embodiments. In some embodiments, the pharmaceutical composition of the present invention can comprise, for example, about 20% of the compound defined in any of the above disclosed embodiments. % to approximately 90%, or 20% to 80%, or 20% to 70%, or 20% to 60%, or 20% to 50%, or 30% to 90%, or 40% to 90%, or 50% to 90%, or 60% to 90%, or 30% to 70%.

[0119]

[0152] In some embodiments, the concentration of the inositol phosphate of the invention (e.g., myo-inositol hexaphosphate or an analog or derivative thereof, or a combination thereof) in each dose of the pharmaceutical composition is about 12.5 mM to about 135 mM. In some versions of this embodiment, the concentration of the inositol phosphate of the invention (e.g., myo-inositol hexaphosphate or an analog or derivative thereof, or a combination thereof) in each dose of the pharmaceutical composition is about 25 mM, about 39 mM, or about 114 mM.

[0120]

[0153] A pharmaceutical composition formulation suitable for parenteral administration comprises a compound defined in any of the above disclosed embodiments mixed with a pharma- ceutically acceptable carrier (such as, for example, sterile water or sterile isotonic saline). Such a formulation can be prepared, packaged, or sold in a form suitable for rapid or continuous administration. Injectable formulations can be prepared, packaged, or sold in unit dosage form, for example, in ampoules or in multi-dose containers containing a preservative. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations. Such formulations can further comprise one or more additional components, including, but not limited to, suspending agents, stabilizing agents, or dispersing agents.

[0121]

[0154] In some embodiments, in formulations for parenteral administration, the active agent (e.g., a compound defined in any of the embodiments disclosed above) is provided in a dry (i.e., powder or granules) form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.

[0122]

[0155] The pharmaceutical composition can be prepared, packaged, or sold in the form of a sterile injectable aqueous or oily suspension or solution. This suspension or solution can be formulated according to known art and can contain, in addition to the active agent (e.g., the compound defined in any of the above-disclosed embodiments), additional ingredients such as dispersing agents, wetting agents, or suspending agents described herein. Such sterile injectable formulations can be prepared using non-toxic parenterally acceptable diluents or solvents, such as water or 1,3-butanediol. Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils, such as synthetic mono- or di-glycerides.

[0123]

[0156] Other parentally administrable formulations that are useful include those that contain the active agent (e.g., a compound defined in any of the embodiments disclosed above) in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer system.

[0124]

[0157] Compositions for sustained release or implantation may include pharma- ceutically acceptable polymeric or hydrophobic materials, such as emulsions, ion exchange resins, sparingly soluble polymers, or sparingly soluble salts.

[0125]

[0158] Controlled or sustained release formulations of the pharmaceutical compositions of the present invention can be made using conventional techniques. In some cases, the dosage form to be used may be prepared using a variety of techniques, such as hydropropyl methylcellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multi-layer coatings, microparticles, liposomes, or other compositions, in varying proportions to provide the desired release profile. The present invention can be applied to the preparation of pharmaceutical compositions of the present invention, such as the pharmaceutical compositions of the present invention, ...

[0126]

[0159] Most controlled-release pharmaceutical products have a common goal of improving therapy compared to that achieved by their non-controlled counterparts. Ideally, the use of an optimally designed controlled-release formulation in medical treatment is characterized by the use of a minimum amount of therapeutic agent to cure or manage a condition in a minimum period of time. Advantages of controlled-release formulations include extended activity of the therapeutic agent, reduced frequency of dosing, and improved patient compliance. In addition, controlled-release formulations can be used to affect the time of onset of action or other characteristics, such as blood levels of the therapeutic agent, and thus the occurrence of side effects.

[0127]

[0160] Most controlled-release formulations are designed to release a certain amount of therapeutic agent initially to rapidly produce the desired therapeutic effect, and then slowly and continuously release another amount of therapeutic agent to maintain this level of therapeutic effect over an extended period of time. In order to maintain this steady level of therapeutic agent in the body, the therapeutic agent must be released from the dosage form at a rate that will replace the amount of therapeutic agent being metabolized or excreted from the body.

[0128]

[0161] The controlled-release of an active agent can be stimulated by various inducers, such as pH, temperature, enzymes, water, or other physiological conditions or compounds. The term "controlled-release component" in the context of the present invention is defined herein as one or more compounds, including but not limited to, polymers, polymer matrices, gels, permeable membranes, liposomes, or microspheres, or combinations thereof, that facilitate the controlled-release of an active agent.

[0129]

[0162] In some embodiments, the formulations of the present invention can be, but are not limited to, short term, fast elimination, as well as controlled release, including sustained release, delayed release and pulsatile release formulations.

[0130]

[0163] The term sustained release is used in its conventional sense to mean a therapeutic agent formulation (e.g., a compound defined in any of the above disclosed embodiments) that provides for gradual release of a therapeutically effective agent over an extended period of time, and that can, but is not necessarily, result in a substantially constant blood level of the therapeutic agent over an extended period of time, which period can be as long as a month or more, and should be a longer release than if the same amount of agent were administered in the form of a bolus dose.

[0131]

[0164] For sustained release, the compound can be formulated with suitable polymers or hydrophobic materials that provide the compound with sustained release properties.Therefore, the compound for use in the method of the present invention can be administered in the form of microparticles, for example, in the form of wafers or disks by injection or by implantation.In some embodiments, the compound of the present invention is administered to a patient alone or in combination with another pharmaceutical agent using sustained release formulations.

[0132]

[0165] The term delayed release is used herein in its conventional sense to mean a therapeutic formulation that provides for an initial release of a therapeutic agent after some delay after administration of the therapeutic agent. The delay can be from about 10 minutes to about 12 hours. The term pulsatile release refers to a therapeutic agent formulation that releases a therapeutic agent in such a manner as to result in a pulsatile plasma profile of the therapeutic agent after administration. The term immediate release is used herein in its conventional sense to mean a therapeutic formulation that provides for release of the therapeutic agent immediately after administration.

[0133]

[0166] Additional formulations and dosage forms of the compositions of the present invention include those disclosed in US6340475, US6488962, US6451808, US5972389, US5582837, and US5007790; US20030147952, 20030104062, 20030104053, 20030044466, 20030039688, and 20020051820; WO2003035041, WO200303504 0, WO2003035029, WO200335177, WO2003035039, WO2002096404, WO2002032416, WO2001097783, WO2001056544, WO2001032217, WO1998055107, WO1998011879, WO1997047285, WO1993018755, and WO1990011757.

[0134]

[0167] The pharmaceutical preparations according to the invention are prepared by methods known in the art, in particular by conventional mixing, coating, granulation, dissolving or lyophilization processes.

[0168] The present invention also provides a compound, a combination of compounds, or a pharmaceutical formulation as defined in any of the above aspects of the invention, given in its broadest definition or as specified in any of the aspects set out above for use as a pharmaceutical.

[0135] 4. Method and route of administration

[0169] In some embodiments, the compound, pharmaceutical composition or combination formulation defined in any of the above disclosed embodiments is administered together, simultaneously or sequentially with another therapeutic agent. In some versions of this embodiment, the additional therapeutic agent comprises cilostazol, pentoxifylline or a combination thereof.

[0136]

[0170] In some embodiments, the compound, pharmaceutical composition or combination preparation defined in any of the above embodiments is provided for administration of an effective amount.The compound, pharmaceutical composition or combination preparation can be administered parenterally, for example, intravenously, intraperitoneally, intramuscularly, intraarterially, intradermally, intrathecally, epidurally or spinally or subcutaneously.Parenteral administration can be by bolus injection or by intravenous infusion.

[0137]

[0171] In a detailed embodiment of the invention, myo-inositol hexaphosphate (or a formulation containing myo-inositol hexaphosphate, such as SNF472) is administered by intravenous infusion. In another detailed embodiment of the invention, myo-inositol hexaphosphate is administered subcutaneously. In another embodiment, a derivative of inositol or a myo-inositol hexaphosphate derivative, such as a derivative of inositol ... 13 is H, n is an integer from 2 to 200, and R1, R5, R9 and R 11 is any group of formula II (or a sodium salt thereof) is administered by intravenous infusion. In another embodiment, a compound of formula I having R3 and R7 = a group of formula V, 13 is H, n is an integer from 2 to 200, and R1, R5, R9 and R 11 is any group of formula II (or a sodium salt thereof), is administered subcutaneously.

[0138]

[0172] Alternatively, the compounds, pharmaceutical compositions or combined preparations can be administered as a component of a hemodialysis, hemofiltration, or peritoneal dialysis solution or system.

[0173] In the particular case of patients treated by dialysis, a very suitable method of administration is to administer the inositol phosphates of the invention (e.g., non-bolus) through a dialysis machine (pre- or post-filtration) instead of injecting the inositol phosphates of the invention directly into the patient's vein. Thus, blood can be treated with an inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) when circulated through a dialysis circuit while the patient is on dialysis, and when blood containing the inositol phosphates of the invention is returned to the body.

[0139]

[0174] Thus, in some embodiments, the compound, pharmaceutical composition or combined preparation defined in any of the above disclosed embodiments is administered to a patient during hemodialysis. In some embodiments, the compound, pharmaceutical composition or combined preparation defined in any of the above disclosed embodiments is administered to the blood extracted from the patient during hemodialysis, preferably before it is filtered (i.e., the therapeutic agent is administered to the unfiltered blood of the patient in the dialysis circuit). In some embodiments, the compound is an inositol hexaphosphate, in particular myo-inositol hexaphosphate sodium salt or a derivative thereof, i.e., R3 and R7=groups of formula V, where R 13 is H, n is an integer from 2 to 200, and R1, R5, R9 and R 11 is a compound of formula I having all groups of formula II (or sodium salts thereof).

[0140]

[0175] In the case of dialysis patients, administration of the inositol phosphates of the present invention (e.g., myo-inositol hexaphosphate) by the dialysis machine allows the blood to equilibrate with the dialysis fluid before being returned to the body; thus, the inositol phosphates of the present invention (e.g., myo-inositol hexaphosphate) can scavenge ionized calcium, a fact that is compensated for when the blood passes through a dialysis filter, thereby eliminating said side effects and significantly improving the safety profile. Furthermore, administration of the inositol phosphates of the present invention (e.g., myo-inositol hexaphosphate) in conjunction with hemodialysis allows the dosage of the compound to be reduced and adverse side effects to be minimized, which is advantageous in terms of reduced toxicity, especially when administered to unfiltered blood extracted from the patient during hemodialysis.

[0141]

[0176] In some embodiments, the compound, pharmaceutical composition or combined formulation defined in any of the above disclosed embodiments is administered to a patient being treated with hemodialysis before or after the dialysis treatment.

[0142]

[0177] In general, the effective dosage of the inositol phosphates of the invention (e.g., myo-inositol hexaphosphate) administered according to the methods disclosed herein will depend, for example, on the relative efficacy of the compounds involved, the severity of the disorder being treated, and the species and weight of the subject. In some embodiments, the effective dosage of the inositol phosphates of the invention for a subject of some species (e.g., human) can be calculated based on experimental data available for a different or reference species (e.g., rat).

[0143]

[0178] Thus, for example, a dose of inositol myo-hexaphosphate administered as part of a regimen involving administration to a rat subject at a dose of 20 mg / kg would be equivalent to administration of the same effective agent to a human subject at a dose of 4.2 mg / kg (i.e., a total dose of 300 mg of inositol myo-hexaphosphate to a human subject weighing approximately 70 kg). Similarly, a dose of 40 mg / kg to a rat subject would be equivalent to administration of inositol myo-hexaphosphate at a dose of 8.4 mg / kg to a human subject as defined above. Doses can be adjusted based on the subject's age, species, body weight, body surface area, renal clearance, sex, pathological condition, route of administration, co-administration of one or more other drugs, and various physiological and psychological factors using methods known in the art (Pan S. et al., Patient Prefer Adherence 2016;10:549-560; Pai M, Pharmacotherapy 2012;32:856-868; Hacker M. et al., eds., Pharmacology: Prin Ciples and Practice (Academic Press; Burlington, MA, USA, 2009). As used herein, the term "mg / kg" means mg of inositol phosphate of the present invention per kilogram of body mass (weight) of the subject.

[0144]

[0179] In some embodiments, the dose of the inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) comprises from about 0.001 mg / kg to about 60 mg / kg of an inositol phosphate, inositol phosphate analog, inositol phosphate derivative, or combination thereof according to the invention. In some further embodiments, the dose of the inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) is from about 0.001 mg / kg to about 20.0 mg / kg, or from about 20.0 mg / kg to about 40.0 mg / kg, or from about 40.0 mg / kg to about 60.0 mg / kg.

[0145]

[0180] In some embodiments, the dose of an inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) is between about 0.001 mg / kg and about 1.0 mg / kg, between about 1.0 mg / kg and about 10.0 mg / kg, between about 10.0 mg / kg and about 20.0 mg / kg, between about 20.0 mg / kg and about 30.0 mg / kg, between about 30.0 mg / kg and about 40.0 mg / kg, between about 40.0 mg / kg and about 50.0 mg / kg, or between about 50.0 mg / kg and about 60.0 mg / kg.

[0146]

[0181] In some embodiments, the dose of an inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) is about 0.001 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.0 mg / kg, about 1.0 mg / kg to about 5.0 mg / kg, about 5.0 mg / kg to about 10.0 mg / kg, about 10.0 mg / kg to about 15.0 mg / kg, about 15.0 mg / kg to about 20.0 mg / kg, about 20.0 mg / kg to about 25.0 mg / kg, about 25.0 mg / kg to about 30.0 mg / kg, about 30.0 mg / kg to about 35.0 mg / kg, about 35.0 mg / kg to about 40.0 mg / kg, about 40.0 mg / kg to about 45.0 mg / kg, or about 45.0 mg / kg to about 50.0 mg / kg.

[0147]

[0182] In some embodiments, the dose of an inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) is about 0.001 mg / kg to about 0.25 mg / kg, about 0.25 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 0.75 mg / kg, about 0.75 mg / kg to about 1.0 mg / kg, about 1.0 mg / kg to about 2.50 mg / kg, about 2.50 mg / kg to about 3.5 mg / kg, about 3.5 mg / kg to about 4.5 mg / kg, about 4.5 mg / kg to about 5.5 mg / kg, about 5.5 mg / kg to about 6.5 mg / kg, about 6.5 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 8.5 mg / kg, about 8.5 mg / kg to about 9.5 mg / kg, about 9.5 mg / kg to about 10.5 mg / kg, about 10.5 mg / kg to about 11.5 mg / kg, about 10.5 mg / kg to about 12.5 mg / kg, about 12.5 mg / kg to about 13.5 mg / kg, about 13.5 mg / kg to about 14.5 mg / kg, about 14.5 mg / kg to about 15.5 mg / kg, about 15.5 mg / kg to about 20.5 mg / kg, about 15.5 mg / kg to about 21.5 mg / kg, about 15.5 mg / kg to about 22.5 mg / kg, about 15.5 mg / kg to about 23.5 mg / kg, about 15.5 mg / kg to about 24.5 mg / kg, about 15.5 mg / kg to about 25.5 mg / kg, about 15.5 mg / kg to about 26 about 5.0 mg / kg to about 5.0 mg / kg, about 5.0 mg / kg to about 7.5 mg / kg, about 7.5 mg / kg to about 10.0 mg / kg, about 10.0 mg / kg to about 12.5 mg / kg, about 12.5 mg / kg to about 15.0 mg / kg, about 15.0 mg / kg to about 17.5 mg / kg, about 17.5 mg / kg to about 20.0 mg / kg, about 20.0 mg / kg to about 22.5 mg / kg, about 22.5 mg / kg to about 25.0 mg / kg, about 25.0 mg / kg to about 30.0 mg / kg, about 25.0 mg / kg to about 35.0 mg / kg, about 25.0 mg / kg to about 36.5 mg / kg, about 25.0 mg / kg to about 37.5 mg / kg, about 25.0 mg / kg to about 38.5 mg / kg, about 25.0 mg / kg to about 39.5 mg / kg, about 39.5 mg / kg to about 40.0 mg / kg, about 40.0 mg / kg to about 42.5 mg / kg, about 40.0 mg / kg to about 44.5 mg / kg, about 44.5 mg / kg to about 45.5 mg / kg, about 45.5 mg / kg to about 50.0 mg / kg, about 45.5 mg / kg to about 52.5 mg / kg, about 45.5 mg / kg to about 56.5 mg / kg, about 45.5 mg / kg to about 58.5 mg / kg, about 45.5 mg / kg to about 59.5 mg / kg, about 59.5 mg / kg to about 60.0 mg / kg, about 59.5 mg / kg to about 61.5 .5mg / kg to about 25.0mg / kg, about 25.0mg / kg to about 27.5mg / kg, about 27.5mg / kg to about 30.0mg / kg, about 30.0mg / kg to about 32.5mg / kg, about 32.5mg / kg to about 35.0mg / kg, about 35.0mg / kg to about 37.5mg / kg, about 37.5mg / kg to about 40.0mg / kg, about 40.0mg / kg to about 42. 5 mg / kg, about 42.5 mg / kg to about 45.0 mg / kg, about 45.0 mg / kg to about 47.5 mg / kg, about 47.5 mg / kg to about 50.0 mg / kg, about 50.0 mg / kg to about 52.5 mg / kg, about 52.5 mg / kg to about 55.0 mg / kg, about 55.0 mg / kg to about 57.5 mg / kg, or about 57.5 mg / kg to about 60.0 mg / kg.

[0148]

[0183] In some embodiments, the dose of an inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) is about 0.25 mg / kg to about 60.0 mg / kg, about 0.5 mg / kg to about 60.0 mg / kg, about 0.75 mg / kg to about 60.0 mg / kg, About 1.0 mg / kg to about 60.0 mg / kg, about 2.50 mg / kg to about 60.0 mg / kg, about 5.0 mg / kg to about 60.0 mg / kg, about 7.5 mg / kg to about 60.0 mg / kg, about 10.0 mg / kg to about 60.0 mg / kg, about 12.5 mg / kg to about 60.0 mg / kg, about 15.0 mg / kg to about 60.0 mg / kg, about 17.5 mg / kg to about 60.0 mg / kg, about 20.0 mg / kg to about 60.0 mg / kg, about 22.5 mg / kg to about 60.0 mg / kg, about 25.0 mg / kg to about 60.0 mg / kg, about 27.5 mg / kg to about 60.0 mg / kg, about 30. 0 mg / kg to about 60.0 mg / kg, about 32.5 mg / kg to about 60.0 mg / kg, about 35.0 mg / kg to about 60.0 mg / kg, about 37.5 mg / kg to about 60.0 mg / kg, about 40.0 mg / kg to about 60.0 mg / kg, about 42.5 mg / kg to about 60.0 mg / kg, about 45.0 mg / kg to about 60.0 mg / kg, about 47.5 mg / kg to about 60.0 mg / kg, about 50.0 mg / kg to about 60.0 mg / kg, about 52.5 mg / kg to about 60.0 mg / kg, about 55.0 mg / kg to about 60.0 mg / kg, or about 57.5 mg / kg to about 60.0 mg / kg.

[0149]

[0184] In some embodiments, the dose of an inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) is from about 0.001 mg / kg to about 57.5 mg / kg, from about 0.001 mg / kg to about 55.0 mg / kg, from about 0.001 mg / kg to about 52.5 mg / kg, from about 0.001 mg / kg to about 50.0 mg / kg, from about 0.001 mg / kg to about 47.5 mg / kg, from about 0.001 mg / kg to about 50.0 ... .001mg / kg ~ approx. 45.0mg / kg, approx. 0.001mg / kg ~ approx. 42.5mg / kg, approx. 0.001mg / kg ~ approx. 40.0mg / kg, approx. 0.001mg / kg ~ approx. 37.5 mg / kg, about 0.001mg / kg to about 35.0mg / kg, about 0.001mg / kg to about 32.5mg / kg, about 0.001mg / kg to about 30.0mg / kg, about 0.001mg / k g ~ approx. 27.5 mg / kg, approx. 0.001 mg / kg ~ approx. 25.0 mg / kg, approx. 0.001 mg / kg ~ approx. 22.5 mg / kg, approx. 0.001 mg / kg ~ approx. 20.0 mg / kg, approx. 0 .001mg / kg ~ approx. 27.5mg / kg, approx. 0.001mg / kg ~ approx. 25.0mg / kg, approx. 0.001mg / kg ~ approx. 22.5mg / kg, approx. 0.001mg / kg ~ approx. 20.0m g / kg, about 0.001 mg / kg to about 17.5 mg / kg, about 0.001 mg / kg to about 15.0 mg / kg, about 0.001 mg / kg to about 12.5 mg / kg, about 0.001 mg / kg to about 10.0 mg / kg, about 0.001 mg / kg to about 7.5 mg / kg, about 0.001 mg / kg to about 5.0 mg / kg, or about 0.001 mg / kg to about 2.5 mg / kg.

[0150] 5. Instructions

[0185] The compounds, pharmaceutical compositions, combined formulations, methods and routes of administration defined in any of the above disclosed embodiments can be used to increase tissue perfusion and / or oxygenation in a subject in need thereof.

[0151]

[0186] As used herein, the term "ischemia-related disease or condition" refers to any disease or condition that is related to or results from ischemic events or damage. Examples of ischemia-related disease or condition include, but are not limited to, cerebrovascular (e.g., stroke, transient ischemic attack (TIA), subarachnoid hemorrhage, vascular dementia), cardiovascular (e.g., myocardial infarction, angina pectoris), gastrointestinal (e.g., colitis), peripheral (e.g., acute limb ischemia) and skin (e.g., cyanosis, gangrene) disease or condition.

[0152]

[0187] In some embodiments, the compounds, pharmaceutical compositions, combination formulations, methods and routes of administration of the present invention can be used for the treatment or prevention of ischemia and / or ischemia-related diseases or conditions in a subject in need thereof.

[0153]

[0188] As used herein, the term "renal failure" refers to a decrease in glomerular filtration rate (GFR) or is a disease that causes progressive loss of kidney function with a concomitant decrease in the index. Renal failure is also known as renal decline or kidney disease. Renal disease can be classified as (i) acute kidney injury (AKI), which is a progressive loss of kidney function that generally causes oliguria and fluid and electrolyte abnormalities, and (ii) chronic kidney disease (CKD), which is a much more gradual loss of kidney function over months or years. Depending on the degree of renal function, five stages of CKD are defined based on GFR as follows: (a) stage 1, normal or high GFR (>90 ml / min); (b) stage 2: mild CKD, GFR=60-89 ml / min; (c) stage 3, moderate CKD, GFR=30-59 m / min; (d) stage 4, severe CKD, GFR=15-29 ml / min; and (e) stage 5, end-stage CKD, GFR<15 ml / min. At stage 5, dialysis or a kidney transplant is necessary to maintain health. AKI and CKD can occur simultaneously, known as acute exacerbation of chronic renal failure.

[0154]

[0189] In some embodiments, the compounds, pharmaceutical compositions, combination formulations, methods and routes of administration of the present invention can be used to increase tissue perfusion in subjects with renal disease. The renal disease in the subject can be acute, chronic or both. In some embodiments, the subject is undergoing dialysis (e.g., peritoneal dialysis, hemodialysis). In further aspects of this embodiment, the subject is undergoing hemodialysis. In some other embodiments, the subject is not undergoing dialysis (e.g., subjects with CKD in stages 1-4). In one version of this embodiment, the subject is administered an inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention at an effective dose of about 0.001 mg / kg to about 60 mg / kg.

[0155]

[0190] In some embodiments, the compounds, pharmaceutical compositions, combination formulations, methods and routes of administration of the present invention can be used to treat or prevent ischemia and / or ischemia-related diseases or conditions in subjects with renal disease. The renal disease in the subject can be acute, chronic, or both. In one version of this embodiment, the subject is undergoing dialysis (e.g., peritoneal dialysis, hemodialysis). In a further form of this version, the subject is undergoing hemodialysis. In another version of this embodiment, the subject is not undergoing dialysis (e.g., a subject with CKD in stages 1-4). In one version of this embodiment, the subject is administered an inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention at an effective dose of about 0.001 mg / kg to about 60 mg / kg.

[0156]

[0191] In some embodiments, the compounds, pharmaceutical compositions, combination formulations, methods and routes of administration of the present invention can be used to improve walking ability in a subject in need thereof. In some embodiments, the compounds, pharmaceutical compositions, methods and routes of administration of the present invention can be used to increase maximal walking distance (MWD), maximal walking time (MWT), or both in a subject in need thereof. In some embodiments, the subject suffers from renal disease. The renal disease in the subject can be acute, chronic, or both. In one version of this embodiment, the subject is undergoing dialysis (e.g., peritoneal dialysis, hemodialysis). In a further form of this version, the subject is undergoing hemodialysis. In another version of this embodiment, the subject is not undergoing dialysis (e.g., a subject with CKD in stages 1-4). In one version of this embodiment, the subject is administered an inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention at an effective dose of about 0.001 mg / kg to about 60 mg / kg.

[0157]

[0192] The compounds, pharmaceutical compositions, combination preparations, methods of administration and routes of administration of the present invention are particularly useful for increasing tissue perfusion and / or oxygenation in the lower limbs, in particular for the treatment and prevention of peripheral arterial disease. A further condition that can benefit from the use of the inositol phosphates of the present invention is critical limb ischemia. In a detailed embodiment, the compounds, pharmaceutical compositions, combination preparations, methods of administration and routes of administration as defined in any of the above disclosed embodiments are The route is for use in increasing tissue perfusion and / or oxygenation, particularly in the lower limbs, and particularly for the treatment and prevention of PAD and / or CLI. In some embodiments, the subject is undergoing dialysis (e.g., peritoneal dialysis, hemodialysis). In some further embodiments, the subject is undergoing hemodialysis. In some other embodiments, the subject is not undergoing dialysis (e.g., a subject with CKD in stages 1-4). In one version of this embodiment, the subject is administered an inositol phosphate of the invention (e.g., myo-inositol hexaphosphate) at an effective dose of about 0.001 mg / kg to about 60 mg / kg.

[0158]

[0193] The following embodiments further illustrate the scope of the invention.

[0194] Embodiment 1. A compound of general formula I, or a pharma- ceutically acceptable salt thereof, for use in increasing tissue perfusion and / or oxygenation in a subject in need thereof.

[0159] [ka]

[0160]

[0195] [In the formula, R 1、 R 3、 R 5、 R 7、 R9 and R 11 is represented by OH, Formulas II, III, IV, V, VI and VII:

[0161] [ka]

[0162]

[0196] (In the formula, n is an integer ranging from 2 to 200, and R 13 is H, methyl, ethyl and C3-C 10 alkyl);

[0197] however:

[0198] R1, R3, R5, R7, R9 and R 11 at least one of is selected from groups of formulae II, III and IV,

[0199] R1, R3, R5, R7, R9 and R 11 are selected from groups of formulae V, VI and VII.

[0163]

[0200] Embodiment 2. A compound for use according to embodiment 1, which is for the treatment or prevention of peripheral arterial disease.

[0201] Embodiment 3. A compound for use according to embodiment 1 or 2, which is for the treatment or prevention of critical limb ischemia.

[0164]

[0202] Embodiment 4. A compound for use according to any of embodiments 1 to 3, for treating a subject undergoing dialysis, preferably hemodialysis.

[0203] Embodiment 5. A compound for use according to any of embodiments 1-4, which is a sodium salt.

[0165]

[0204] Embodiment 6: R1, R3, R5, R7, R9 and R 11 wherein at least two, at least three, at least four, at least five or at least six of the groups are selected from groups of formulae V, VI and VII.

[0166]

[0205] Embodiment 7.R 1、 R 3、 R 5、 R 7、 R9 and R 11 At least two of , at least 3, at least 4, at least 5 or at least 6 are groups of formula V.

[0167]

[0206] Embodiment 8. A compound for use according to embodiment 7, wherein the compound of formula I is inositol hexaphosphate.

[0207] Embodiment 9. A compound for use according to embodiment 8, which is a hexasodium salt.

[0168]

[0208] Embodiment 10.

[0209] R7, OSO3 - where R1, R3, R5, R9 and R 11 But OPO3 2- , OPSO2 2- or OSO3 - are independently selected from;

[0210] R9, R5 and R1 are OPO3 2- and R7, R3 and R 11 But OSO3 - and;

[0211] R9, R5 and R1 are OSO3 - and R7, R3 and R 11 But OPO3 2- and;

[0212] R3, R1 and R 11 But OSO3 - R9, R7 and R5 are OPO3 2- and;

[0213] R3, R1 and R 11 But OPO3 2- R9, R7 and R5 are - and;

[0214] R7 and R1 are OPO3 2- and R9, R5, R3, and R 11 But OPO3 - and;

[0215] R7 and R1 are OSO3 - and R9, R5, R3, and R 11 But OPO3 2- and;

[0216] R7 and R5 are OPO3 2- and R9, R3, R1, and R 11 But OSO3 - or

[0217] R7 and R5 are OSO3 - and R9, R3, R1, and R 11 But OPO3 2- The compound for use according to embodiment 7, wherein

[0169]

[0218] Embodiment 11. A compound for use according to any of embodiments 1 to 10, wherein the compound of formula I has a myo-inositol conformation.

[0219] Embodiment 12. R1, R3, R5, R7, R9 and R 11The compound for use according to any of embodiments 1 to 6, wherein 0, 1, 2 or 3 of are selected from groups of formulae V, VI and VII.

[0170]

[0220] Embodiment 13. R1, R3, R5, R7, R9 and R 11 four of which are groups of formula II, R1, R3, R5, R7, R9 and R 11 and n is 0 or 1. The compound for use according to embodiment 12, wherein two of are selected from groups of formulae V, VI and VII.

[0171]

[0221] Embodiment 14. R1, R3, R5, R7, R9 and R 11 four of which are groups of formula II, R1, R3, R5, R7, R9 and R 11 and n is a radical of formula V.

[0172]

[0222] EMBODIMENT 15. (i) R1, R5, R9 and R 11 is a group of formula II, R3 and R7 are selected from groups of formula V, VI and VII, (i) R1, R3, R9 and R 11 A compound for use according to any of embodiments 12 to 14, wherein is a group of formula II, and R5 and R7 are selected from groups of formula V, VI and VII.

[0173]

[0223] Embodiment 16. A compound for use according to embodiment 15, wherein the group selected from V, VI and VII is a group of formula V.

[0224] Embodiment 17. A compound for use according to any of embodiments 12-16, wherein the group of formula V, VI or VI has n in the range of 2 to 200.

[0174]

[0225] Embodiment 18. A compound for use according to embodiment 17, wherein n is in the range of 9 to 30.

[0226] Embodiment 19. A compound for use according to embodiment 18, wherein n is in the range of 15 to 30.

[0175]

[0227] Embodiment 20. A compound for use according to embodiment 17, wherein n is in the range of 3 to 9.

[0228] Embodiment 21.R 13 The compound for use according to any of embodiments 12-20, wherein

[0176]

[0229] Embodiment 22. R1, R5, R9 and R 11 A compound for use according to embodiment 21, wherein R is a group of formula II and R3 and R7 are groups of formula V.

[0230] Embodiment 23. A pharmaceutical composition for use as defined in any of embodiments 1 to 4, comprising a compound as defined in any of embodiments 1 to 22, together with pharma- ceutically acceptable excipients and carriers.

[0177]

[0231] Embodiment 24. A pharmaceutical composition according to embodiment 23, wherein the compound is present in an amount of 20-90% (w / w) of the total composition.

[0232] Embodiment 25. A pharmaceutical composition according to embodiment 24, wherein the compound is present in an amount of 30-80% (w / w) of the total composition.

[0178]

[0233] Embodiment 26. A pharmaceutical composition according to embodiment 25, wherein the compound is present in an amount of 40-70% (w / w) of the total composition.

[0234] Embodiment 27. A pharmaceutical composition according to any of embodiments 23-26, in a dry form for reconstitution with a suitable vehicle.

[0179]

[0235] Embodiment 28. A pharmaceutical composition according to any of embodiments 23 to 26, which is a solution, preferably an isotonic saline solution.

[0236] Embodiment 29. A pharmaceutical composition according to any of embodiments 23 to 27 forming part of a hemodialysis, hemofiltration, or peritoneal dialysis solution.

[0180]

[0237] Embodiment 30. A pharmaceutical composition according to any of embodiments 23 to 29, which is for controlled release.

[0238] Embodiment 31. A compound for use according to any one of embodiments 1 to 22 or a pharmaceutical composition for use according to any one of embodiments 23 to 30, which is administered to a patient undergoing dialysis.

[0181]

[0239] Embodiment 32. A compound for use according to embodiment 31, wherein the dialysis is hemodialysis.

[0240] Embodiment 33. A compound or pharmaceutical composition for use according to embodiment 31 or 32, which is administered before dialysis.

[0182]

[0241] Embodiment 34. A compound or pharmaceutical composition for use according to embodiment 31 or 32, which is administered during dialysis.

[0242] Embodiment 35. A compound or pharmaceutical composition for use according to embodiment 31 or 32, which is administered after dialysis.

[0183]

[0243] Embodiment 36. A compound or pharmaceutical composition for use according to any of embodiments 31 to 35, which is administered by parenteral route.

[0244] Embodiment 37. A compound or pharmaceutical composition for use according to embodiment 36, wherein parenteral administration is intravenous, subcutaneous or intramuscular.

[0184]

[0245] Embodiment 38. A compound or pharmaceutical composition for use according to embodiment 37, wherein intravenous administration is by bolus administration or by intravenous infusion.

[0246] Embodiment 39. A compound or pharmaceutical composition for use according to embodiment 34, administered to unfiltered blood extracted from a patient.

[0185]

[0247] Embodiment 40. A compound or pharmaceutical composition for use according to embodiments 1-39, wherein the compound is administered to the subject in a therapeutically effective amount of from about 0.001 mg / kg to about 60 mg / kg.

[0248] Embodiment 41. A compound or pharmaceutical composition for use according to embodiment 40, wherein the compound is administered to the subject in a therapeutically effective amount of about 15 mg / kg to about 45 mg / kg.

[0186]

[0249] Embodiment 42. (i) A combination preparation comprising (a) at least one compound according to any one of embodiments 1 to 22 or (b) at least one pharmaceutical composition according to any one of embodiments 23 to 30, and (ii) at least one additional therapeutic agent for use in human health.

[0187]

[0250] Embodiment 43. A combination formulation according to embodiment 42, wherein the additional therapeutic agent is cilostazol, pentoxifylline or a combination thereof.

[0251] Embodiment 44. A method for increasing tissue perfusion and / or oxygenation in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition according to any one of embodiments 1 to 41 or a combination formulation according to embodiment 42 or 43.

[0188]

[0252] Embodiment 45. A method for treating or preventing ischemia and / or ischemia-related diseases or conditions in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition according to any one of embodiments 1 to 41 or a combination formulation according to embodiment 42 or 43.

[0189]

[0253] Embodiment 46. A method for improving walking ability in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition according to any one of embodiments 1 to 41 or a combination formulation according to embodiment 42 or 43.

[0190]

[0254] Embodiment 47. A method for increasing maximal walking distance (MWD), maximal walking time (MWT), or both, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition according to any one of embodiments 1 to 41, or a combination formulation according to embodiment 42 or 43.

[0191]

[0255] Embodiment 48. A method for treating or preventing peripheral arterial disease in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound or pharmaceutical composition according to any one of embodiments 1 to 41 or a combination formulation according to embodiment 42 or 43.

[0192]

[0256] Embodiment 49. The method according to any one of embodiments 44 to 48, wherein the combined preparations are administered to the subject together, simultaneously or sequentially.

[0257] The present invention is further illustrated by the following examples, which should not be construed as limiting. The contents of all references cited throughout this application are hereby incorporated by reference in their entirety.

[0193] General Procedure 1. Limb Blood Perfusion

[0258] Position limb blood perfusion and ischemic status (i.e., blood perfusion including perfusion units, perfusion difference, and perfusion ratio) are assessed by laser Doppler perfusion imaging using a PeriCam PSI NR imager (Perimed AB, Jarfalla, SE). Subjects are anesthetized with 3% isoflurane delivered in 100% oxygen at a flow rate of 1 L / min prior to measurements. Perfusion difference and perfusion ratio are calculated by comparing baseline and any indicated intermediate or final measurements for each group. Blood flow is measured using C-values ​​of the active agents being tested (i.e., 15 min after treatment with SNF472, 20 min after treatment with IP4-BIS-PEG100, and 3-4 h after treatment with cilostazol). max Evaluate around.

[0194] 2. Walking ability test

[0259] Quantification of ambulatory performance (maximal walking time (MWT) and maximum walking distance (MWD)) is measured by a forced incremental treadmill running test. A 2-lane rodent treadmill (LE8709TS; PanLab / Harvard Apparatus, Holliston, MA, US) is used. The treadmill is maintained at a speed of 15 m / min (25 cm / sec) for the first 5 minutes at 15% incline running, then at 33 cm / sec for the next 5 minutes, and finally at a speed of 40 cm / sec for up to an additional 30 minutes.

[0195]

[0260] Limb function was assessed by the C-values ​​of the active agents being tested (i.e., 15 min after treatment with SNF472 and 3–4 h after treatment with cilostazol). max Subjects are assessed at 0.05-0.05°C for acclimatization on a treadmill according to a set protocol prior to testing. Subjects who do not comply with the protocol are excluded from the study.

[0196]

[0261] Subjects are allowed to continue running for up to 40 minutes during testing or until exhaustion (i.e., subjects remain on the shock grid for 5 consecutive seconds). MWD and MWT are then calculated for each animal.

[0197] 3. Tissue and Blood Collection, Calcification Assay

[0262] Subjects are anesthetized by isoflurane inhalation. Blood is obtained by exsanguination through cardiac puncture. Subjects are then sacrificed and their tissues (e.g., right and left femoral arteries, aorta) are harvested. Blood and tissues are processed and analyzed to determine their calcium content.

[0198]

[0263] Calcium content in tissue samples was determined using Optima® according to the manufacturer's instructions. Quantification is performed by inductively coupled plasma optical emission spectroscopy (ICP-OES) using a 7300 DV ICP-OES System spectrometer (PerkinElmer, Inc., Waltham, MA, US). Plasma myo-inositol-hexaphosphate levels are quantified by LC-MS / MS chromatographic methods described in the art. See WO2013050603.

[0199] Example 1 Prevention of Limb Ischemia Effects of hemoperfusion

[0264] SNF472, IP4-4,6-bisPEG100 sodium salt and Cilostazol The preventive effect of zole on blood perfusion was tested using a rat model for a period of 12 days. Limb ischemia in subjects was induced from D1, except for the sham group, in which ischemia was not induced. The ischemia-induced subjects were then treated with placebo and active drug combinations from D1 to D12 to evaluate their impact in preventing limb ischemia. Observations were made at several points during treatment from D1 to D12. All subjects were weighed daily before treatment.

[0200] 1. Induction of Limb Ischemia

[0265] Fifty-four male Sprague-Dawley (SD) rats (Envigo Corp., Huntingdon, GB), weighing approximately 250-275 g, were used. Subjects were fed the AO4 diet (Scientific Animal Food & Engineering; Carpe Bio, Amersfoort, NL). Subjects were divided into five groups with 8-10 rats per group as follows:

[0201]

[0266] Group 1 - Control (sham)

[0267] Group 2a: Placebo - saline

[0268] Group 2b: Placebo - 5% carboxymethylcellulose (CMC) sodium salt solution

[0269] Group 3 - SNF472(Na6IP6)

[0270] Group 4 - IP4-4,6-bisPEG100 sodium salt

[0271] Group 5 - Cilostazol (C 20 H 27 N5O2)

[0272] In subjects in groups 2a-5, limb ischemia was induced by subcutaneous administration of 120,000 IU / kg of vitamin D3 (cholecalciferol, dufafural D3 1000; Zoetis Inc., Parsippany, NJ, US) in saline (2 mL / kg) daily from D1 to D3. Subjects in sham group 1 received saline without vitamin D3 (2 mL / kg) subcutaneously daily from D1 to D3.

[0202]

[0273] Subjects in groups 1 and 2a received saline (2 mL / kg) subcutaneously every day from D1 to D12. Subjects in group 2b received 5% CMC sodium salt in aqueous solution (5 mL / kg) orally every day from D1 to D12.

[0203]

[0274] Administration of Vitamin D3 induced ischemia in the hind limbs of subjects in Groups 2a, 2b, and 3-5 was assessed by laser Doppler perfusion imaging. 2. Effects on Ischemia Rescue and Limb Blood Perfusion – Laser Doppler Imaging Assay

[0275] In subjects of groups 3 and 4, limb hemoperfusion was induced by subcutaneous administration of SNF472 (Na6IP6; free base: 600 g / mol) 20 mg / kg and IP4-4,6-bisPEG100 sodium salt (free base 696.27 g / mol), respectively, in saline (2 mL / kg) daily from D1 to D12. In subjects of group 5, cilostazol (CMC) was administered in 5% sodium salt in aqueous solution (5 mL / kg) daily from D1 to D12. 20 H 27 Limb hemoperfusion was induced by oral administration of 20 mg / kg N5O2, free base 369.46 g / mol; Lot No. LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US).

[0204]

[0276] Limb ischemia status was assessed in all rats by laser Doppler perfusion imaging on D0 (baseline), D6, and D12. Perfusion difference and perfusion ratio were calculated by comparing either the D0 baseline or D6 and D12 measurements for each group. In particular, perfusion difference and perfusion ratio were calculated by comparing measurements for group 1 (control) and (a) 2a and 2b placebo groups (i.e., saline, 5% CMC sodium salt in aqueous solution), (b) group 3 (SNF472), (c) group 4 (IP4-4,6-bisPEG100 sodium salt), and (d) group 5 (cilostazol). The calculation was made by:

[0205]

[0277] On the days when hemoperfusion studies with active drugs were performed (ie, D6, D12), doses were administered as follows:

[0278] Group 3 - SNF472 (Na6IP6): 15 minutes before measurement

[0279] Group 4 - IP4-4,6-bisPEG100 sodium salt: 20 minutes before measurement

[0280] Group 5 - Cilostazol (C 20 H27 N5O2): 3 to 4 hours before measurement

[0281] Under the above scheme, effective drugs are those that have the highest serum concentration (C max ) should be.

[0206]

[0282] Subjects in Group 1 did not show any significant changes in their perfusion parameters.

[0283] Administration of SNF472 and IP4-4,6-bisPEG100 sodium salt attenuated the decline in blood perfusion in the limbs of subjects in groups 3-5 (i.e., treated with vitamin D3) compared to placebo and cilostazol. These results suggest that SNF472 and IP4-4,6-bisPEG100 are more effective than cilostazol in increasing blood perfusion in the hind limbs of treated subjects. See Figures 7, 8, and 9.

[0207] 3. Calcium Content and Mineralization – ICP-OES

[0284] SNF472 was shown to be effective against vascular calcification, as it inhibited aortic calcification (31±16%) after daily subcutaneous administration of 20 mg / kg compared to placebo. Cilostazol was not active against calcification at the same dose. See Figure 10.

[0208] 4. Pharmacokinetics

[0285] Subjects from groups 1, 2, 3 and 5 were sacrificed after exsanguination. They were then autopsied and their aortas were harvested. The tissues were freeze-dried for 24 h and weighed. The freeze-dried tissues were then digested with a 1:1 HNO3:HClO4 mixture at 180 °C for 2-4 h in a dry bath incubator. The digested tissues were subsequently diluted with ultra-pure Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US) to a final volume of 10 mL. Calcium content in the aortic samples was quantified by ICP-OES.

[0209]

[0286] Subjects from groups 1, 2, 3, 4 and 5 were anesthetized and their blood obtained on D12. Approximately 8-10 mL of whole blood was collected per subject and separated into blood collection tubes for plasma (K3EDTA, approximately 6 mL of blood) and serum (approximately 2-3 mL of blood) laboratory testing. Plasma was stored in one aliquot of 600 μL and several other aliquots of 500 μL were stored. Serum was separated into two aliquots.

[0210]

[0287] Plasma myo-inositol-hexaphosphate levels (i.e., C 15 min after the final SNF472 dose) in subjects from Group 3 max The amount of IgG1-related ...

[0211]

[0288] All subjects in group 3 were fully exposed to the SNF472 product. Plasma levels 15 minutes after the last subcutaneous dose on D12 were 15587±ng / mL (24±12 μM). Plasma levels in rats after daily dosing at 20 mg / kg were comparable to those found in hemodialysis patients treated with SNF472 by the intravenous route at a dose of 4.2 mg / kg.

[0212] Example 2 Prevention of Limb Ischemia Effects on maximal walking (MWT) and distance (MDT) times

[0289] The preventive effects of SNF472 and cilostazol on blood perfusion, walking ability and tissue calcification were tested using a rat model for 24 days. In addition, the effect of combined treatment of SNF472 and cilostazol on blood perfusion was also tested. Limb ischemia was induced in subjects from D1 to D3, except for the sham group, in which ischemia was not induced. The ischemia-induced subjects were then treated with placebo and active drug combinations from D1 to D12 to evaluate their effects in preventing (a) limb ischemia and tissue calcification and (b) deterioration of walking ability. Treatment in all subjects was discontinued from D13 to D24. Observations were made at several points during and after the treatment phase from D1 to D24. All subjects were weighed daily before treatment.

[0213] 1. Induction of Limb Ischemia

[0290] Fifty-four male Sprague-Dawley (SD) rats (Envigo Corp., Huntingdon, GB), weighing approximately 250-275 g, were used. Subjects were fed the AO4 diet (Scientific Animal Food & Engineering; Carpe Bio, Amersfoort, NL). Subjects were divided into five groups, with 8-10 animals per group, as follows:

[0214]

[0291] Group 1 - Control (sham)

[0292] Group 2a: Placebo - saline

[0293] Group 2b: Placebo - 5% CMC sodium salt solution

[0294] Group 3 - SNF472(Na6IP6)

[0295] Group 4 - Cilostazol (C 20 H 27 N5O2)

[0296] Group 5 - Cilostazol + SNF472

[0297] In subjects in groups 2a-5, limb ischemia was induced by subcutaneous administration of 120,000 IU / kg of vitamin D3 (cholecalciferol, dufafural D3 1000; Zoetis Inc., Parsippany, NJ, US) in saline (2 mL / kg) daily from D1 to D3. Subjects in sham group 1 received subcutaneous administration of saline (2 mL / kg) without vitamin D3 daily from D1 to D3.

[0215]

[0298] Subjects in groups 1 and 2a received saline (2 mL / kg) subcutaneously every day from D1 to D12. Subjects in group 2b received 5% CMC sodium salt in aqueous solution (5 mL / kg) orally every day from D1 to D12.

[0216] 2. Effects on Ischemia Rescue and Blood Perfusion – Laser Doppler Imaging Assay

[0299] Subjects in groups 1 and 2a received 2 mL / kg of saline by subcutaneous route daily from D1 to D12. Subjects in group 2b received 5 mL / kg of 5% CMC sodium salt in aqueous solution orally daily from D1 to D12. Furthermore, subjects in group 3 received 20 mg / kg of SNF472 (Na6IP6, free base: 600 g / mol) sodium salt (free base 696.27 g / mol) in saline (2 mL / kg) by subcutaneous route daily from D1 to D12. Subjects in group 4 received 5 mL / kg of cilostazol (C6IP6, free base: 600 g / mol) in aqueous solution (5 mL / kg) daily from D1 to D12. 20 H 27 Subjects in Group 5 were orally administered 20 mg / kg of N5O2, free base 369.46 g / mol; Lot No. LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US). 20 H 27 N5O2, free base 369.46 g / mol; Lot No. LRAB9590, Sigma-Aldrich Corp., St. Louis (ii) 20 mg / kg of SNF472 (Na6IP6, free base: 600 g / mol) in saline (2 mL / kg) was administered by subcutaneous route daily from D1 to D12.

[0217]

[0300] On days when hemoperfusion studies with active drugs were performed (ie, D6, D12, and D18), doses were administered as follows:

[0301] Group 3 - SNF472 (Na6IP6): 15 minutes before measurement

[0302] Group 4 - Cilostazol (C 20 H 27 N5O2) 3 to 4 hours before measurement

[0303] Group 5 - Cilostazol + SNF472: 15 minutes before the test for SNF472 and 3-4 hours before the test for Cilostazol

[0304] Under the above scheme, active drugs are those that are at their maximum plasma concentration (C max ) should be.

[0218]

[0305] Limb ischemia status was assessed by laser Doppler perfusion imaging in all rats during and after treatment and interruption (i.e., D0, D6, D12, D18). Perfusion difference and perfusion ratio were calculated by comparing the baseline and any measurements at D6, D12, and D18 for each group. In particular, perfusion difference and perfusion ratio were calculated by comparing the measurements of group 1 (control) and (a) 2a and 2b placebo groups (saline, 5% CMC sodium salt solution), (b) 3 (SNF472), (c) 4 (cilostazol), and (d) 5 (cilostazol / SNF472 combination).

[0219]

[0306] SNF472 alone or in combination with cilostazol attenuated limb ischemia in rats. Treatment with cilostazol alone was not effective in this model. The SNF472 effect on blood perfusion was maintained every 6 days after treatment was discontinued. See Figure 11.

[0220] 3. Effects on walking ability – treadmill running test

[0307] Maximal walking time (MWT) and maximum walking distance (MWD) were assessed in groups 1-4 on D0, D5, D10, and D17 by treadmill running test (8-10 animals per group and time point).

[0221]

[0308] Subjects were allowed to acclimate to the treadmill for 2 days before testing. On the first day, subjects were exercised for 5-10 minutes at treadmill speeds ranging from 15 m / min to 24 m / min in a progressive manner. On the second day, subjects were first exercised for 5 minutes at a speed of 15 m / min. Subjects were then exercised for an additional 5 minutes at 19.8 m / min (33 cm / sec). Finally, subjects were exercised for up to an additional 30 minutes at 24 m / min (40 cm / sec). Preoperative walking time and distance records were obtained. Animals that did not follow the protocol were excluded from the study.

[0222]

[0309] Limb function (MWT and MWD) was assessed using a treadmill running test 15 min after the corresponding daily administration in groups 1, 2, and 3. Limb function in group 4 was assessed 3-4 h after treatment.

[0223]

[0310] Under the above scheme, active drugs are those that have their maximum plasma concentration (C max ) should be.

[0311] Subjects were allowed to continue running for 40 min or until exhaustion (i.e., subjects remained on the shock grid for 5 consecutive seconds). MWD and MWT were then calculated for each animal.

[0224]

[0312] SNF472 and cilostazol improved walking ability in rats compared to vehicle (+54% MWD and +46% MWT). See Figure 13. Moreover, the effect of SNF472 on improving walking ability was maintained even 5 days after treatment was discontinued (D17). In contrast, cilostazol lost its beneficial effect immediately after treatment was stopped. See Figure 13.

[0225] 4. Calcium Content and Mineralization – ICP-OES

[0313] Subjects were anesthetized and their blood was obtained on D24. Subjects were sacrificed after exsanguination. They were then autopsied and their aortas were harvested. Tissues were freeze-dried for 24 h and weighed. The freeze-dried tissues were then digested with a 1:1 HNO3:HClO4 mixture at 180 °C for 2-4 h in a dry bath incubator. The digested tissues were subsequently rehydrated with ultrapure Milli-Q water (MilliporeSigma, Merck). The tissue samples were diluted to a final volume of 10 mL with 10% KGaA (Burlington, MA, US). Calcium content in the tissue samples was quantified by ICP-OES.

[0226]

[0314] SNF472 was shown to be effective against vascular calcification, as it inhibited aortic calcification (41±9%) after daily subcutaneous administration at 20 mg / kg compared to placebo. Cilostazol was not active against calcification at the same dose. See Figure 14.

[0227] Example 3 Treatment of Limb Ischemia

[0315] After the initiation of limb ischemia, the effects of SNF472 and cilostazol on blood perfusion, walking ability and tissue calcification were tested in a rat model for a period of 13 days. Limb ischemia was induced in all groups from D1 to D3, except for the sham group, where ischemia was not induced. The subjects with induced ischemia were administered placebo and active drug combinations from D5 onwards to generate ischemia before the start of treatment. From D5 to D13, subjects were treated to evaluate the effects of treatment on limb ischemia, walking ability and tissue calcification. Observations were made at several points during the treatment from D1 to D13. All subjects were weighed daily before treatment.

[0228] 1. Induction of Limb Ischemia

[0316] Sixty-six male Sprague-Dawley (SD) rats (Envigo Corp., Huntingdon, GB), weighing approximately 250-275 g, were used. Subjects were fed the AO4 diet (Scientific Animal Food & Engineering; Carpe Bio, Amersfoort, NL). Subjects were divided into five groups with 8-14 animals per group as follows:

[0229]

[0317] Group 1 - Control (sham)

[0318] Group 2 – D5 Ca baseline

[0319] Group 3a: Placebo - saline

[0320] Group 3b: Placebo - 5% CMC sodium salt solution

[0321] Group 4 - SNF472(Na6IP6)

[0322] Group 5 - Cilostazol (C 20 H 27 N5O2)

[0323] Limb ischemia was induced by subcutaneous administration of 120,000 IU / kg of vitamin D3 (cholecalciferol, dufafural D3 1000; Zoetis Inc., Parsippany, NJ, US) in saline (2 mL / kg) daily from D1 to D3 in subjects in groups 2 to 5. Subjects in group 1 received subcutaneous administration of saline (2 mL / kg) without vitamin D3 daily from D1 to D3.

[0230] 2. Effects on Ischemia Rescue and Blood Perfusion – Laser Doppler Imaging Assay

[0324] Subjects in group 1 received 2 mL / kg of saline subcutaneously every day from D1 to D13. Subjects in group 2 received 2 mL / kg saline by subcutaneous route daily from D1 to D5. On D5, 4 subjects in group 1 and all subjects in group 2 were sacrificed to determine their Ca baseline values.

[0231]

[0325] Subjects in placebo group 3a received 2 mL / kg of saline by subcutaneous route daily from D5 to D13. Subjects in placebo group 3b received 5 mL / kg of 5% CMC sodium salt in aqueous solution orally daily from D5 to D13. Additionally, subjects in group 4 received 40 mg / kg of SNF472 (Na6IP6, free base: 600 g / mol) in saline (2 mL / kg) by subcutaneous route daily from D5 to D13.

[0232]

[0326] Subjects in group 5 were administered cilostazol (CMC) in 5% sodium salt in aqueous solution (5 mL / kg). 20 H 27 N5O2, free base 369.46 g / mol; Lot No. LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) 40 mg / kg was orally administered daily from D5 to D13. The 40 mg / kg dose of cilostazol in rats is comparable to the therapeutic dose of 8.4 mg / kg in PAD patients.

[0233]

[0327] On the days when hemoperfusion studies with active drugs were performed (ie, D5, D13), doses were administered as follows:

[0328] Group 4 - SNF472 (Na6IP6): 15 minutes before measurement

[0329] Group 5 - Cilostazol (C 20 H 27 N5O2): 3 to 4 hours before measurement

[0330] Under the above scheme, active drugs are those that are at their maximum plasma concentration (C max ) should be.

[0234]

[0331] Limb function and ischemic status were assessed by laser Doppler perfusion imaging in all groups on D0 and D5, and in groups 1, 3a, 3b, 4 and 5 on D13. Perfusion difference and perfusion ratio were calculated by comparing the baseline and either D5 or D13 measurements for each group. In particular, perfusion difference and perfusion ratio were calculated by comparing the measurements for group 1 (control) and (a) 3a and 3b placebo groups (saline, 5% CMC sodium salt solution), (b) group 4 (SNF472), (c) group 5 (cilostazol).

[0235]

[0332] VitD3 administration induces a drop in blood perfusion in hind limbs (measured on D5, immediately before administering therapy) in groups 3a, 3b, 4 and 5.On D13, only animals treated with SNF472 show significant improvement in limb blood perfusion compared to D5 before treatment.No improvement or ischemic rescue is reported on D13 in animals treated with placebo or cilostazol compared to D5.See Figure 16.

[0236] 3. Effects on walking ability – treadmill running test

[0333] Maximum walking time (MWT) and maximum walking distance (MWD) were assessed by treadmill running test (8-12 animals per group and time point) in all groups on D0 and in groups 1, 3a, 3b, 4, and 5 on D6 and D11. Subjects were acclimated to the treadmill for 2 days before testing. On the first day, subjects were exercised for 5-10 minutes at treadmill speeds ranging progressively from 15 m / min to 24 m / min. On the second day, subjects were initially exercised for 5 minutes at a speed of 15 m / min. Subjects were then exercised for an additional 5 minutes at 19.8 m / min (33 cm / sec). Finally, subjects were exercised for up to an additional 30 minutes at 24 m / min (40 cm / sec). Preoperative walking time and distance records were obtained. Animals that did not follow the protocol were excluded from the study.

[0237]

[0334] Limb function (MWT and MWD) was measured using a treadmill 15 min after the corresponding daily administration. The linear running test was used to evaluate limb function in groups 1, 3a, 3b, and 4. Limb function in group 5 was evaluated 3-4 hours after treatment.

[0238]

[0335] Under the above scheme, active drugs are those that are at their maximum plasma concentration (C max ) should be.

[0336] Subjects were allowed to continue running for 40 min or until exhaustion (i.e., subjects remained on the shock grid for 5 consecutive seconds). MWD and MWT were then calculated for each animal.

[0239]

[0337] Even when treatment was started 5 days after induction of ischemia, SNF472 improved the walking ability of rats compared to vehicle (+49% MWD), whereas cilostazol was not effective under the same conditions and at therapeutic doses (40 mg / kg / day). See Figure 16.

[0240] 4. Calcium Content and Mineralization – ICP-OES

[0338] Four subjects in group 1 and all subjects in group 2 were sacrificed on D5 to determine their Ca baseline values. The remaining subjects in group 1 and all subjects in groups 3a, 3b, 4, 5, and 6 were sacrificed on D13.

[0241]

[0339] Subjects were sacrificed after exsanguination. They were then autopsied and their right and left femoral arteries were harvested. Tissues were freeze-dried for 24 h and weighed. The freeze-dried tissues were then digested with a 1:1 HNO3:HClO4 mixture at 180 °C for 2-4 h in a dry bath incubator. The digested tissues were subsequently diluted to a final volume of 10 mL with ultrapure Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US). Calcium content in tissue samples was quantified by ICP-OES.

[0242]

[0340] SNF472 was shown to be effective against vascular calcification on D13. Compared to placebo, SNF472 inhibited calcification in the femoral artery by approximately (30%) after daily subcutaneous administration of 40 mg / kg. See Figure 17.

[0243] 5. Pharmacokinetics

[0341] Subjects from groups 1, 3a, 3b, 4 and 5 were anesthetized and their blood obtained on D13. Approximately 8-10 mL of whole blood was collected per subject and separated into blood collection tubes for plasma (K3EDTA, approximately 6 mL blood) and serum (approximately 2-3 mL blood) laboratory testing. Plasma was stored as one aliquot of 600 μL and several other aliquots of 500 μL. Serum was separated into two aliquots.

[0244]

[0342] Myo-inositol-hexaphosphate levels in plasma of subjects in Group 4 (i.e., C 15 min after the final SNF472 dose) max The amount of IgG1-related ...

[0245]

[0343] All subjects in group 4 were fully exposed to the SNF472 product. Plasma levels 15 minutes after the last subcutaneous dose of D13 were 40078±15024ng / mL (60.7±22.8uM). Plasma levels in rats after daily dosing at 20mg / kg were comparable to those found in hemodialysis patients treated with SNF472 at a dose of 8.4mg / kg by the intravenous route.

[0246] Example 4 SN472 Drug Interactions

[0344] The compatibility of SNF472 with other drugs regularly prescribed to subjects with reduced renal function was analyzed.

[0247]

[0345] Wistar rats (Charles River Labs, Inc., Wilmington, Mass., US) were treated with: (i) SNF472 subcutaneously (sc), (ii) SNF472 + sevelamer (oral), (iii) SNF472 (sc) + cinacalcet (oral), (iv) SNF472 (sc) + Vit D (sc), (v) SNF472 (sc) + sodium thiosulfate (sc), and SNF472 (sc) + ibandronate (sc). No significant differences were observed between administration of SNF472 alone or together with any of the other drugs assayed.

[0248] Example 5 Prevention of Limb Ischemia Dose-response effects of SNF472 on maximal walking (MWT) and distance (MDT) times

[0346] The preventive effects of several different doses of SNF472 and the highest tolerated dose of cilostazol on blood perfusion, walking ability and tissue calcification were tested in a rat model for 12 days. Limb ischemia was induced in subjects from D1 to D3, except for the sham group. The ischemia-induced animals were then treated with placebo and active drug combinations from D1 to D12 to evaluate their impact in preventing (a) limb ischemia and tissue calcification and (b) deterioration of walking ability. Observations were made at several points during the treatment phase from D1 to D12. All subjects were weighed daily before treatment.

[0249] 1. Induction of Limb Ischemia

[0347] One hundred two male Sprague-Dawley (SD) rats (Envigo Corp., Huntingdon, GB), weighing approximately 250-275 g, were used. Subjects were fed the AO4 diet (Scientific Animal Food & Engineering; Carpe Bio, Amersfoort, NL). Subjects were divided into five groups with 8-12 animals per group as follows:

[0250]

[0348] Group 1 - Control (sham)

[0349] Arm 2a Placebo - saline, subcutaneous

[0350] Arm 2b: Placebo - 5% CMC sodium salt solution, oral

[0351] Group 3 - SNF472 (Na6IP6) 1mg / kg, s.c.

[0352] Group 4 - SNF472 (Na6IP6) 7.5 mg / kg, s.c.

[0353] Group 5 - SNF472 (Na6IP6) 15 mg / kg, s.c.

[0354] Group 6 - SNF472 (Na6IP6) 30 mg / kg, s.c.

[0355] Group 7 - SNF472 (Na6IP6) 45 mg / kg, s.c.

[0356] Group 8 - Cilostazol (C20 H 27 N5O2) 45mg / kg, orally

[0357] In subjects in groups 2–8, limb ischemia was induced by subcutaneous administration of 120,000 IU / kg of vitamin D3 (cholecalciferol, dufafural D3 1000; Zoetis Inc., Parsippany, NJ, US) in saline (2 mL / kg) daily from D1 to D3. Subjects in sham group 1 received subcutaneous administration of saline (2 mL / kg) without vitamin D3 daily from D1 to D3.

[0251]

[0358] Subjects in groups 1 and 2a received saline (2 mL / kg) subcutaneously every day from D1 to D12. Subjects in group 2b received 5% CMC sodium salt in aqueous solution (5 mL / kg) orally every day from D1 to D12.

[0252] 2. Effects on Ischemia Rescue and Blood Perfusion – Laser Doppler Imaging Assay

[0359] Subjects in groups 1 and 2a received 2 mL / kg of saline by subcutaneous route daily from D1 to D12. Subjects in group 2b received 5 mL / kg of 5% CMC sodium salt in aqueous solution orally daily from D1 to D12. Additionally, subjects in groups 3, 4, 5, 6 and 7 received 1 mg / kg, 7.5 mg / kg, 15 mg / kg, 30 mg / kg and 45 mg / kg of SNF472 (Na6IP6, free base: 600 g / mol) sodium salt (free base 696.27 g / mol), respectively. SNF472 was administered daily in saline (2 mL / kg) by subcutaneous route from D1 to D12. Subjects in group 8 received cilostazol (C6IP6, free base: 600 g / mol) in aqueous solution (5 mL / kg). 20 H 27 N5O2, free base 369.46 g / mol; Lot No. LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) 45 mg / kg was orally administered daily from D1 to D12.

[0253]

[0360] On the days when hemoperfusion studies with active drugs were performed (ie, D6 and D12), doses were administered as follows:

[0361] Groups 3, 4, 5, 6 and 7 - SNF472 (Na6IP6): 15 minutes before measurement

[0362] Group 8 - Cilostazol (C 20 H 27 N5O2) 3 to 4 hours before measurement

[0363] Under the above scheme, active drugs are those that are at their maximum plasma concentration (C max ) should be.

[0254]

[0364] Limb ischemia status was assessed by laser Doppler perfusion imaging in all rats during the treatment period (i.e., D0, D6 and D12). Perfusion difference and perfusion ratio were calculated by comparing the baseline and either D6 and D12 measurements for each group. In particular, perfusion difference and perfusion ratio were calculated by comparing the measurements of group 1 (control) and (a) 2a and 2b placebo groups (saline, 5% CMC sodium salt solution), (b) 3, 4, 5, 6 and 7 (SNF472) and (c) 8 (cilostazol).

[0255]

[0365] SNF472 attenuated limb ischemia in rats in a dose-response manner. Treatment with cilostazol alone was ineffective in this model. 3. Effects on walking ability – treadmill running test

[0366] Maximal walking time (MWT) and maximum walking distance (MWD) were assessed in groups 1-8 on D0, D5, and D10 by treadmill running test (8-12 animals per group and time point).

[0256]

[0367] Subjects were allowed to acclimate to the treadmill for 2 days before testing. On the first day, subjects were exercised for 5-10 minutes at treadmill speeds ranging from 15 m / min to 24 m / min in a progressive manner. On the second day, subjects were first exercised for 5 minutes at a speed of 15 m / min. Subjects were then exercised for an additional 5 minutes at 19.8 m / min (33 cm / sec). Finally, subjects were exercised for up to an additional 30 minutes at 24 m / min (40 cm / sec). Preoperative walking time and distance records were obtained. Animals that did not follow the protocol were excluded from the study.

[0257]

[0368] Limb function (MWT and MWD) was assessed using a treadmill running test 15 min after the corresponding daily administration in groups 1, 2, 3, 4, 5, 6 and 7. Limb function in group 8 was assessed 3-4 h after treatment.

[0258]

[0369] Under the above scheme, active drugs are those that have their maximum plasma concentration (C max ) should be.

[0370] Subjects were allowed to continue running for 40 min or until exhaustion (i.e., subjects remained on the shock grid for 5 consecutive seconds). MWD and MWT were then calculated for each animal.

[0259]

[0371] SNF472 and cilostazol improved the walking ability in rats compared to vehicle, and the effect of SNF472 on improving the walking ability was dose-response dependent.

[0260] 4. Calcium Content and Mineralization – ICP-OES

[0372] Subjects were anesthetized and their blood was obtained on D12. Subjects were sacrificed after exsanguination. They were then autopsied and their hearts and aorta arteries were harvested. Tissues were freeze-dried for 24 h and weighed. The freeze-dried tissues were then digested with a 1:1 HNO3:HClO4 mixture at 180°C for 2-4 h in a dry bath incubator. The digested tissues were subsequently diluted to a final volume of 10 mL with ultrapure Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US). Calcium content in tissue samples was quantified by ICP-OES.

[0261]

[0373] SNF472 was shown to be effective against cardiac and vascular calcification, including cardiac and aortic arterial calcification, in a dose-response manner. Cilostazol was not active against calcification at the same doses.

[0262] Example 6 Prevention of limb ischemia in an adenine-induced uremic rat model Effects of SNF472 on limb blood perfusion

[0374] The preventive effects of SNF472 and cilostazol doses on blood perfusion and tissue calcification were examined for 21 days in a relevant rat model of chronic kidney disease.

[0263]

[0375] Uremia and limb ischemia in animals were induced from D1 to D21, except for the sham group, in which neither uremia nor ischemia was induced. The ischemia-induced animals were then treated with placebo and active drug combinations from D1 to D21 to evaluate their effects in preventing (a) limb ischemia and (b) tissue calcification. Observations were made at several points during the treatment phase from D1 to D21. All subjects were weighed daily before treatment.

[0264] 1. Induction of uremia and limb ischemia

[0376] Sixty-eight male Sprague-Dawley (SD) rats (Envigo Corp., Huntingdon, GB), weighing approximately 250-275 g, were used and fed a pelleted high-phosphorus diet (SM R, 10 mm chow, 1.06% Ca, 1.03% P) (SSNIFF Spezialdiaeten, Soest, DE) ad libitum. Subjects were divided into six groups with 8-12 animals per group as follows:

[0377] Group 1 - Control (sham)

[0378] Arm 2a: Placebo - saline, subcutaneous, once daily

[0379] Arm 2b: Placebo - saline, subcutaneous, Alzet pump, 4 weeks

[0380] Group 3 - SNF472 (Na6IP6) 30mg / kg, subcutaneously, once a day

[0381] Group 4 - SNF472 (Na6IP6) 45mg / kg, subcutaneously, once daily

[0382] Group 5 - Alzet pumps containing SNF472 (Na6IP6) total dose 400mg / 4 weeks (100mg / week) were implanted subcutaneously on D1 before adenine administration for 4 weeks.

[0265]

[0383] Group 6 - Cilostazol (C 20 H 27 N5O2) 45mg / kg / day, By mouth, once a day

[0384] Uremia and limb ischemia were induced in groups 2-6 by a daily dose of adenine (500 mg / kg suspended in 1% carboxymethylcellulose, administered orally) for the first 10 days, followed by a dose of α-calcidol (100 ng / kg in olive oil, administered orally) three times per week from D11 to D19 to accelerate and homogenize the development of cardiovascular calcification and ischemia.

[0266]

[0385] On day 21, the animals were sacrificed and blood and tissue samples were taken. Serum creatinine (reference number OSR6178) and urea (reference number OSR6134) levels were determined using the corresponding Beckman Coulter assay kits (Beckman Coulter, Inc., Brea, CA, US).

[0267]

[0386] The first sham group animals were orally administered a 1% carboxymethylcellulose solution (5 mL / kg) daily from D1 to D10, followed by olive oil three times per week from D11 to D19. Neither uremia nor ischemia was induced in the sham group.

[0268] 2. Effects on Ischemia Rescue and Blood Perfusion – Laser Doppler Imaging Assay

[0387] Subjects in group 2a received saline (2 mL / kg) subcutaneously twice daily for 21 days. Subjects in group 2b received saline subcutaneously using an Alzet pump for 4 weeks.

[0269]

[0388] Additionally, animals in groups 3 and 4 received 30 and 45 mg / kg of SNF472 (Na6IP6, free base: 600 g / mol) sodium salt (free base 696.27 g / mol) subcutaneously twice daily. SNF472 was administered by the subcutaneous route in saline (2 mL / kg) twice daily from D1 to D21. Animals in group 5 received 400 mg / kg of SNF472 dissolved in saline subcutaneously for 4 weeks using an Alzet pump.

[0270]

[0389] Animals in group 6 were administered cilostazol (CMC) in 5% sodium salt in water (5 mL / kg). 20 H 27 N5O2, free base 369.46 g / mol; Lot No. LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) 45 mg / kg was orally administered daily from D1 to D21.

[0271]

[0390] On days when hemoperfusion studies with active drugs were performed (i.e., D10, D17, and D21), doses were administered as follows:

[0391] Groups 3 and 4 - SNF472 (Na6IP6): 15 minutes before measurement

[0392] Group 6 - Cilostazol (C 20 H 27 N5O2) 3 to 4 hours before measurement

[0393] Under the above scheme, active drugs are those that are at their maximum plasma concentration (C max ) should be.

[0272]

[0394] Limb ischemia status was assessed by laser Doppler perfusion imaging in all rats during treatment (i.e., D0, D10, and D17 and D21). Perfusion difference and perfusion ratio were calculated by comparing baseline and D10, D17, and D21 measurements for each group. In particular, perfusion difference and perfusion ratio were calculated by comparing measurements for group 1 (control) and (a) 2a and 2b placebo groups, (b) 3, 4, and 5 (SNF472), and (c) 6 (cilostazol).

[0273]

[0395] SNF472 reduces rat limb ischemia in uremic rats in a dose-response manner Treatment with cilostazol alone was ineffective in this model. 3. Calcium Content and Mineralization – ICP-OES

[0396] Subjects were anesthetized and their blood was obtained on D21. Subjects were sacrificed after exsanguination. They were then autopsied and their hearts and aortic arteries were harvested. Tissues were freeze-dried for 24 h and weighed. The freeze-dried tissues were then digested with a 1:1 HNO3:HClO4 mixture at 180°C for 2-4 h in a dry bath incubator. The digested tissues were subsequently diluted with ultrapure Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US) to a final volume of 10 mL. Calcium content in tissue samples was quantified by ICP-OES.

[0274]

[0397] SNF472 was shown to be effective against cardiac and vascular calcification in uremic rats in a dose-response manner, e.g., in cardiac and aortic arterial calcification, after daily subcutaneous administration and / or delivery by the Alzet pump, compared with placebo. Cilostazol was not active against calcification at the same dose.

Claims

1. A compound of general formula I, or a pharma- ceutically acceptable salt thereof, for use in increasing tissue perfusion and / or oxygenation in a subject in need thereof. [Case 1] [In the formula, R １、 R ３、 R ５、 R ７、 R ９ and R １１ are independently OH, a group of formula II, III, IV: [Case 2] and heterologous moieties, with the proviso that: (i) R １ , R ３ , R ５ , R ７ , R ９ and R １１ is selected from the groups of formulae II, III and IV, (ii) R １、 R ３、 R ５、 R ７、 R ９ and R １１ provided that 0, 1, 2 or 3 of the

2. A compound of general formula I, or a pharma- ceutically acceptable salt thereof, for use in the treatment or prevention of ischemia and / or an ischemia-related disease or condition in a subject in need thereof. [C3] [In the formula, R １ , R ３ , R ５ , R ７ , R ９ and R １１ are independently OH, a group of formula II, III, IV: [C4] and heterologous moieties, with the proviso that: (i) R １ , R ３ , R ５ , R ７ , R ９ and R １１ is selected from the groups of formulae II, III and IV, (ii) R １、 R ３、 R ５、 R ７、 R ９ and R １１ provided that 0, 1, 2 or 3 of the

3. The heterogeneous moiety is a group of formula V, a group of formula VI, and a group of formula VII: [C5] [In the formula, n is an integer ranging from 2 to 200, R １３ is selected from H, methyl and ethyl.

4. 4. A compound for use according to any one of claims 1 to 3 for the treatment or prevention of peripheral arterial disease.

5. 5. A compound for use according to any one of claims 1 to 4, for the treatment or prevention of critical limb ischemia.

6. 6. The compound for use according to any one of claims 1 to 5, which is a sodium salt.

7. 7. A compound for use according to any one of claims 1 to 6, wherein the compound of formula I is inositol hexaphosphate.

8. 8. The compound for use according to claim 7, wherein the inositol hexaphosphate is myo-inositol hexaphosphate.

9. 9. The compound for use according to claim 7 or 8, which is a hexasodium salt.

10. R １ , R ３ , R ５, R ７ , R ９ and R １１ The compound for use according to any one of claims 1 to 6, wherein one or two of are selected from the groups of formulae V, VI and VII.

11. R １ , R ５ , R ９ and R １１ is a group of formula II, R ３ and R ７ A compound for use according to claim 10, wherein is a group of formula V

12. The group of formula V is where n is in the range of 2 to 200, and R １３ The compound for use according to claim 11, wherein is H.

13. A pharmaceutical composition for use as defined in any one of claims 1, 3 to 5, comprising a compound as defined in any one of claims 1 to 12, together with pharma- ceutically acceptable excipients and carriers.

14. A compound for use according to any one of claims 1 to 12 or a pharmaceutical composition for use according to claim 13, administered to a subject with renal failure.

15. A compound for use according to any one of claims 1 to 12 or a pharmaceutical composition for use according to claim 13, which is administered to a subject undergoing dialysis.

16. 16. The compound or pharmaceutical composition for use according to claim 15, administered to a subject undergoing hemodialysis.

17. 17. The compound or pharmaceutical composition for use according to claim 16, which is administered to unfiltered blood extracted from a subject.

18. A compound for use according to any one of claims 1 to 12 or a pharmaceutical composition for use according to claim 13, which is administered by parenteral route.

19. 19. The compound or pharmaceutical composition for use according to claim 18, wherein parenteral administration is intravenous, subcutaneous or intramuscular.

20. A compound of general formula I, or a pharma- ceutically acceptable salt thereof, as defined in any one of claims 1 to 12, or a pharmaceutical composition as defined in claim 13, for use in improving the walking ability of a subject in need thereof.

21. A compound of general formula I, or a pharma- ceutically acceptable salt thereof, as defined in any one of claims 1 to 12, or a pharmaceutical composition as defined in claim 13, for use in increasing maximal walking distance (MWD), maximal walking time (MWT), or both, in a subject in need thereof.

22. 23. The compound or pharmaceutical composition for use according to any one of claims 1 to 22, wherein the compound is administered to a subject at an effective dose that is from about 0.001 mg / kg to about 60 mg / kg.

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