Inositol phosphate compounds for use in increasing tissue perfusion

Abstract

The present invention relates to inositol phosphates, their analogs, derivatives and pharmaceutically acceptable salts for use in increasing tissue perfusion and / or oxygenation in subjects in need thereof, particularly in peripheral arterial disease. The present invention also relates to pharmaceutical compositions comprising said inositol phosphates, their analogs, derivatives and pharmaceutically acceptable salts, and their use in increasing tissue perfusion and / or oxygenation and for treating and preventing peripheral arterial disease.

Description

[Technical Field]

[0001]

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

[0002] This invention relates to the use of inositol phosphate (IP), its analogues and derivatives, for increasing tissue perfusion and / or oxygen supply. The invention also relates to pharmaceutical compositions comprising said IP, its analogues and derivatives, and their use in animal and human health. [Background technology]

[0002]

[0003] Peripheral artery disease (PAD) is a common disorder characterized by narrowing and / or occlusion of the arteries of the lower extremities, resulting in reduced muscle perfusion and oxygen supply. PAD is 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 a result of ischemia in the lower extremities. Its primary cause is atherosclerosis. In its milder form, PAD may be limited to intermittent claudication and pain in the lower extremities. Lower extremity PAD is a leading 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. Currently, more than 8 million men and women in the United States alone have PAD. It is estimated that more than 200 million individuals worldwide have 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. Compared to the general population, people with PAD have increased cardiovascular (CV) morbidity and mortality, accelerated rate of functional decline, and increased rates of mobility loss.

[0005]

[0006] The objectives of the treatment protocol for patients with PAD include reducing the rate of cardiovascular events, improving functional activity, and preventing functional decline and loss of mobility. Restoring or improving blood perfusion to the extremities may help achieve these goals.

[0006]

[0007] While endovascular and lower extremity revascularization procedures significantly improve walking ability in patients with PAD, revascularization is not a treatment option for many patients due to the presence of co-existing conditions or because the location and type of atherosclerotic disease in the lower extremities makes revascularization unsuitable. Revascularization is invasive, costly, and carries risks, especially in elderly patients. For these reasons, there is a clinician need for effective, accessible, and tolerable medical therapies to improve lower extremity function in patients with PAD.

[0007]

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

[0008]

[0009] Cilostazol is a phosphodiesterase inhibitor that improves treadmill walking ability by approximately 25% to 40% in individuals with symptomatic PAD. Cilostazol is a phosphodiesterase type 3 inhibitor that acts by increasing the intracellular concentration of cyclic adenosine monophosphate. In this process, the drug acts as a direct arterial vasodilator, inhibiting platelet aggregation and improving 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 confusion. There is a black-bordered warning against prescribing cilostazol to patients 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 impaired renal function or cardiovascular disease, such as cinacalcet, clopidogrel, and ibandronate, and therefore increases the risk of adverse reactions in these patients resulting from the concomitant use of cilostazol with other drugs. [Overview of the project] [Problems that the invention aims to solve]

[0010]

[0011] In conclusion, medical therapies for symptom relief are limited, and while surgical or endovascular interventions may be useful in some individuals, the long-term outcomes are often disappointing. Consequently, there is a need to develop new, more effective, and safer therapies for treating PAD. [Means for solving the problem]

[0011]

[0012] In a first aspect, the present invention relates to a compound of general formula I, or a pharmaceutically acceptable salt thereof, for use in increasing tissue perfusion and / or oxygen supply in subjects requiring increased tissue perfusion and / or oxygen supply.

[0012] [ka]

[0013]

[0013] [wherein, R 1、 R 3、 R 5、 R 7、 R9 and R 11 These are independently OH, the base of formulas II, III, and IV:

[0014] [ka]

[0015] and selected from heterologous parts,

[0014] However:

[0015] R1, R3, R5, R7, R9 and R 11 At least one of these is selected from the bases of equations II, III, and IV,

[0016] R1, R3, R5, R7, R9 and R 11 [The condition is that 0, 1, 2, or 3 of these are parts of a heterogeneous structure.] Regarding.

[0016]

[0017] In another aspect, the present invention relates to a compound of general formula I as defined above for use in the treatment or prevention of ischemia in subjects requiring treatment or prevention of ischemia. In a version of this aspect, the present invention relates to a compound of general formula I as described above for use in the treatment or prevention of ischemia-related diseases or conditions in subjects requiring treatment or prevention of such diseases or conditions.

[0017]

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

[0018] [ka]

[0019] Selected from,

[0019] n is an integer in the range of 2 to 200, R 13 This is selected from H, methyl, or ethyl.

[0020]

[0020] In a further embodiment, the present invention also relates to a method for increasing tissue perfusion and / or oxygen supply, comprising the step of administering a therapeutically effective amount of the compound of formula I, as defined above, together with a pharmaceutically acceptable excipient or carrier, to a subject that requires an increase in tissue perfusion and / or oxygen supply. This embodiment can also be formulated as the use of the compound of formula I, as defined above, for the manufacture of a pharmaceutical for increasing tissue perfusion and / or oxygen supply in a subject that requires an increase in tissue perfusion and / or oxygen supply.

[0021]

[0021] In another embodiment, the present invention also relates to a method for treating or preventing ischemia and / or ischemia-related diseases or conditions, comprising the step of administering a therapeutically effective amount of the compound of formula I, as defined above, together with a pharmaceutically acceptable excipient or carrier, to a subject that is in need of treatment or prevention of ischemia and / or ischemia-related diseases or conditions. This embodiment can also be formulated as the use of the compound of formula I, as defined above, for the manufacture of a pharmaceutical product for treating or preventing ischemia and / or ischemia-related diseases or conditions in a subject that is in need of treatment or prevention of ischemia and / or ischemia-related diseases or conditions.

[0022]

[0022] In a further embodiment, the present invention relates to a method for treating or preventing peripheral artery disease, comprising the step of administering a therapeutically effective amount of the compound of formula I, as defined above, together with a pharmaceutically acceptable excipient or carrier, to a subject in need of treating or preventing peripheral artery disease. This embodiment can also be formulated as the use of the compound of formula I, as defined above, for the manufacture of a pharmaceutical product for treating or preventing peripheral artery disease in a subject in need of treating or preventing peripheral artery disease.

[0023]

[0023] The compounds of the present invention are particularly useful for increasing tissue perfusion and / or oxygen supply in the lower extremities, and are especially useful for the treatment or prevention of peripheral artery 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 reference drug currently shown 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 tissue perfusion and / or oxygen supply, (ii) treating or preventing ischemia and / or ischemia-related diseases, and / or (iii) treating or preventing PAD in subjects requiring treatment or prevention of PAD. 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 pharmaceutical for (i) increasing tissue perfusion and / or oxygen supply, (ii) treating or preventing ischemia and / or ischemia-related diseases, and / or (iii) treating or preventing PAD in subjects requiring treatment or prevention of PAD. [Brief explanation of the drawing]

[0025] [Figure 1]

[0025] This figure shows a representative example of an inositol phosphate analog in which two of the six X molecules are OPSO2 2- and the remaining X molecule is OSO3. It shows two specific forms: 4,6-di-(O-thiophosphate)-inositol-1,2,3,5-tetra-O-sulfate. [Figure 2]

[0026] This figure shows inositol phosphate analogs and inositol phosphate derivatives that can be used to carry out the methods of the present invention. The molecules shown are myo-inositol-pentaphosphate-2-PEG400, myo-inositol hexakissulfate, and silo-mio-inositol hexakissulfate. [Figure 3]

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

[0028] This figure shows exemplary inositol phosphate analogs and inositol phosphate derivatives that can be used to carry out the method of the present invention. R1 represents a heterogeneous structural portion (e.g., PEG or PG). n can be 2 to 200. [Figure 5]

[0029] This figure shows exemplary inositol phosphate analogs and inositol phosphate derivatives that can be used to carry out the method of the present invention. n can be 2 to 200. [Figure 6]

[0030] This figure shows exemplary inositol phosphate analogs and inositol phosphate derivatives that can be used to carry out the method of the present invention. n can be 2 to 200. [Figure 7]

[0031] This figure shows blood flow in the hind limb of a rat model at D0, measured by Doppler laser imaging. Blood flow is shown in normalized perfusion units (PU). Normalization is achieved by comparing the raw data at D0 with the first group data. [Figure 8]

[0032] This figure shows blood flow in the hind limb of a rat model at D6, measured by Doppler laser imaging. Blood flow is shown in normalized perfusion units (PU). Normalization is achieved by comparing the raw data at D6 with the first group data. [Figure 9]

[0033] This figure shows blood flow in the hind limb of a rat model at D12, measured by Doppler laser imaging. Blood flow is shown in normalized perfusion units (PU). Normalization is achieved by comparing the raw data at D12 with the first group data. [Figure 10]

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

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

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

[0037] This figure shows the maximum walking distance (MWD) in a rat model at D17 (5 days after treatment discontinuation), as measured by a treadmill test. The maximum walking distance is shown in meters (m) up to a walking time of 40 minutes. [Figure 14]

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

[0039] This figure 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 achieved by comparing the raw data at D0, D5, and D13 with the first group data. [Figure 16]

[0040] This figure 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), as measured by a treadmill test. The maximum walking distance is shown in meters (m), and the maximum walking time is shown in minutes (min). [Figure 17]

[0041] This figure shows the percentage of suppression of femoral artery calcification in a VitD rat model at D13 (9 days after the start of treatment). Calcium levels at the time of sacrifice were measured by ICP-OES. [Modes for carrying out the invention]

[0026]

[0042] The present invention provides compounds, pharmaceutical compositions, methods of administration and routes for use in increasing tissue perfusion and / or oxygen supply. The present invention also provides compounds, pharmaceutical compositions, methods of administration and routes 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 oxygen supply in the lower extremities, especially for the treatment or prevention of peripheral artery 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 present invention includes embodiments in which only one member of the group exists, is used with, or is related to, a given product or process. The present invention also includes embodiments in which two or more or all of the group members exist, is used with, or is related to, a given product or process.

[0029]

[0045] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this description relates. 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) provide general dictionaries containing many of the terms used herein.

[0030]

[0046] Units, prefixes, and symbols are shown in the form recognized by their International System of Units (SI). Numerical ranges encompass the numbers that define the range. When ranges of values ​​are enumerated, it is understood that each intervening integer value between the enumerated upper and lower bounds of that range, and each fraction thereof, are also disclosed in detail along with each partial range between such values. The upper and lower bounds of any range can independently be included in or excluded from that range, and each range that includes either limit, does not include either limit, or includes both limits is also included within the scope of this invention.

[0031]

[0047] Where values ​​are explicitly enumerated, it should be understood that values ​​having approximately the same content or quantity as the enumerated values ​​are also within the scope of the invention. Where combinations are disclosed, each subcombination of the elements of that combination is also disclosed in detail and is within the scope of the invention. Conversely, where different elements or groups of elements are disclosed individually, their combinations are also disclosed. Where any element of an invention is disclosed as having multiple alternatives to that invention, and each alternative is also disclosed herein, either individually excluded or in any combination with other alternatives; where two or more elements of an invention may have such exclusions, and all combinations of elements having such exclusions are disclosed herein.

[0032]

[0048] As used herein, the term "and / or" is interpreted as each specific invention of a component that includes or does not include two identified features or other characteristics. Accordingly, as used herein in the form of a phrase such as "A and / or B," the term "and / or" is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Similarly, as used in the form of a phrase such as "A, B, and / or C," the term "and / or" is intended to include each of the following embodiments, namely 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 (alone); B (alone); and C (alone).

[0033]

[0049] As used herein and as applied to one or more values ​​of interest, the term “about” means a value similar to the given reference value. In some aspects, the term “about” means a value within the range of 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction of the given reference value (greater than or less than that value), unless otherwise stated or evident from the context (unless such figure exceeds 100% of a possible value).

[0034]

[0050] As used herein, the terms “critical limb ischemia” or “CLI” mean severe occlusion of an artery that significantly reduces blood flow to the limb and severe pain, further progressing to skin ulcers, tenderness, or gangrene. Critical limb ischemia is a very severe condition of peripheral artery disease. In some embodiments, administration of inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention to subjects in need improves their ability to walk faster and longer distances compared to untreated cases.

[0035]

[0051] As used herein, the term “compound” means including all isomers and isotopes of the illustrated structure. 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 may exist as stereoisomers, e.g., double bond isomers (i.e., geometric E / Z isomers) or diastereomers (i.e., 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 mixtures of enantiomers and stereoisomers, e.g., racemates. Enantiomers and stereomeric mixtures of compounds, as well as means for separating them into their constituent enantiomers or stereoisomers, are well known. The compounds, salts, or complexes of the present invention can be prepared in combination with a solvent or water molecule to form solvated compounds and hydrates by routine methods.

[0036]

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

[0037]

Chemical Formula

[0038] .

[0053] As used herein, with respect to (i) a compound of general formula I (e.g., inositol phosphate, an inositol phosphate analog, an inositol phosphate derivative, or a combination thereof), or (ii) a pharmaceutical composition comprising at least one compound of item (i), the term "effective amount", and related terms "effective dosage" and "effective dose", refer to an amount sufficient to produce a beneficial or desired effect. In some embodiments, the beneficial or desired outcome is, for example, a clinical outcome, and thus "effective amount" depends on the context in which it is applied. In the context of administering a therapeutic agent that increases tissue perfusion and / or oxygen supply, an effective amount of the therapeutic agent is, for example, an amount sufficient to (a) enhance tissue perfusion in a specified region, (b) stop, reduce, slow the progression of, or reverse ischemia in a specified region, or (c) improve mobility or walking ability (e.g., speed, distance) in a subject, as compared to the same parameter 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 of blood supply to tissues, which results in a deficiency of oxygen required to maintain cellular metabolism. Ischemia includes not only insufficient oxygen, but also decreased availability of nutrients and inadequate removal of metabolic waste. Ischemia can be partial (reduced perfusion) or complete.

[0040]

[0055] As used herein, the terms “maximum walking distance” or “MWD” mean the distance at which a subject is unable to continue walking on their own due to exhaustion or severe pain. With regard to evaluating the increase in MWD, such increase is evaluated by comparing the subject’s MWD value before and after treatment with the therapeutic agent, or by comparing the MWD value of a control group that was not treated with the therapeutic agent with the MWD value of the control group after treatment.

[0041]

[0056] As used herein, the terms “maximum walking time” or “MWT” mean the time at which a subject is unable to continue walking on their own due to exhaustion or severe pain. With regard to evaluating an increase in MWT, such increase is evaluated by comparing the subject’s MWT value before and after treatment with the therapeutic agent, or by comparing the MWT value of a control group that was not treated with the therapeutic agent with the MWT value of the control group after treatment.

[0042]

[0057] As used herein, the terms “parenteral administration” and “administered parenterally” typically mean modes of administration other than enteral and topical administration, usually by injection, and without limitation include intravenous, intramuscular, intra-arterial, subarachnoid, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions (e.g., renal dialysis infusions).

[0043]

[0058] As used herein, the term “peripheral artery disease” or “PAD” means narrowing of the peripheral blood vessels to the legs (most commonly), stomach, arms, and head. Symptoms include intermittent claudication (e.g., leg pain during walking that improves 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” mean to suppress the onset of a disease or condition in a subject or to reduce its occurrence (for example, to avoid the development of ischemic tissue in a limb).

[0045]

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

[0046]

[0061] As used herein, the terms “subject,” “individual,” “animal,” or “mammal” mean any subject, in particular a mammal subject for which diagnosis, prognosis, or therapy is desired. Mammal subjects include, but are not limited to, humans, general livestock, farm animals, zoo animals, sport animals, companion animals, e.g., dogs, cats, guinea pigs, rabbits, rats, mice, horses, cattle, cows; primates, e.g., apes, monkeys, orangutans, and chimpanzees; canids, e.g., dogs and wolves; felids, e.g., cats, lions, and tigers; equine, e.g., horses, donkeys, and zebras; bears; food animals, e.g., cows, pigs, and sheep; ungulates, e.g., deer and giraffes; rodents, e.g., mice, rats, hamsters, and guinea pigs; and so on. In some embodiments, the subject is a human subject. In some embodiments, the subjects are human patients with reduced tissue perfusion and / or oxygen supply in the muscles of the lower extremities or at risk of developing such a condition. In some further embodiments, the subjects are human patients with ischemia and / or ischemia-related disease or condition, or at risk of developing such ischemia, ischemia-related disease or condition.

[0047]

[0062] As used herein, the term “substantially” means a qualitative condition indicating a complete or near-complete range or degree of the feature or characteristic of interest. Those skilled in the biological art will understand that it is rare, if any, for biological and chemical phenomena to proceed to completion and / or toward completeness or to reach or avoid an absolute outcome. Therefore, the term “substantially” is used herein to capture the inherent potential lack of completeness in many biological and chemical phenomena.

[0048]

[0063] As used herein, the term “tissue perfusion” means 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 area in a subject after administration of the inositol phosphate of the present invention, compared to the same parameters observed in the subject before administration of the therapeutic agent or in a control population that did not receive the therapeutic agent.

[0049]

[0064] As used herein, the terms “to treat” or “to cure” mean the administration of a compound or pharmaceutical composition of the present invention to (i) slow progression, (ii) inhibit progression, (iii) halt the progression of a disease or condition, or (iv) reverse the progression of a disease or condition, after the appearance of its clinical signs. Control of disease progression is understood to mean beneficial or desired clinical outcomes, including, but not limited to, symptom relief, reduction of disease duration, stabilization of the pathological condition (especially to avoid further exacerbations), delay of disease progression, improvement of the pathological condition, and remission (partial and complete). Control of disease progression also includes an extension of survival time compared to the predicted survival time if treatment had not been applied. In the context of the present invention, the terms “to treat” or “to heal” mean, in more detail, (a) to increase tissue perfusion and / or oxygen supply, or (b) to stop, reduce, slow down or reverse the development of ischemic tissue in the lower extremities, in particular, or (c) to improve mobility or walking ability (e.g., speed, distance, endurance) in a subject administered with the compounds or pharmaceutical compositions of the present invention.

[0050]

[0065] As used herein, the term "walking ability" means the ability of an object to move autonomously without assistance. The parameters MWD and MWT indicate the object's walking ability.

[0051] 2.Compound

[0066] The compounds for use in the present invention are inositol phosphates, as well as their analogs and derivatives, as defined in the first embodiment of the present invention. As used herein, the term “inositol phosphate” means a compound having an inositol ring and one, two, three, four, five or six 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 is homogeneous, 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 is homogeneous, e.g., IP6). In some embodiments, the inositol phosphate is a mixture containing, for example, variable amounts of IP1, IP2, IP3, IP4, IP5, and IP6. In some embodiments, inositol phosphate is a racemic mixture.

[0052]

[0067] The present invention also envisions inositol phosphate analogues. As used herein, “inositol phosphate analogue” means a compound having a ring with a different number of carbon atoms relative to the inositol ring (i.e., 5 or 7 carbon atoms) and / or having at least one sulfate or thiophosphate group. For example, a compound comprising a ring with 5, 6, or 7 carbon atoms and at least one phosphate, sulfate, or thiophosphate group should be considered an inositol phosphate analogue.

[0053]

[0068] As used herein, the term “inositol phosphate derivative” means inositol phosphate or inositol phosphate analogues, which contain a heterostructured moiety (i.e., a group that is not a phosphate, sulfate, or thiophosphate). For example, inositol pentasulfate containing a polyethylene glycol heterostructured moiety, or myo-inositol hexaphosphate containing a polyglycerin heterostructured moiety, should be considered inositol phosphate derivatives.

[0054]

[0069] As used herein, the term “heterogeneous structural moiety” means a group in a compound of formula I that is not a phosphate, sulfate, or thiophosphate, but which imparts a desired property to such compound. For example, a heterogeneous structural moiety (e.g., polyglycerin or polyethylene glycol) can increase the solubility of a compound. In some embodiments, a heterogeneous structural moiety can impart several 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 Invention” and “Inositol Phosphate of the Invention” are collective terms encompassing “Inositol Phosphate,” “Inositol Phosphate Analogues,” “Inositol Phosphate Derivatives,” and combinations thereof. In some embodiments, the term “Inositol Phosphate of the Invention” encompasses compositions comprising “Inositol Phosphate,” “Inositol Phosphate Analogues,” 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 present invention that include a ring having 5, 6, or 7 carbon atoms and at least one sulfate or thiophosphate group, but lacking a phosphate group, should be considered "inositol phosphate analogs" or "inositol phosphate analogs" in the context of the present invention. Therefore, the term "inositol phosphate of the present invention" includes not only phosphate-containing compounds but also phosphate-lacking compounds that include a ring having 5, 6, or 7 carbon atoms and at least one sulfate or thiophosphate 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 myo-stereostructure form. In addition to myo-inositol, other naturally occurring stereoisomers of inositol are silo-, muco-, 1D-chiro-, 1L-chiro-, neo-inositol, allo-, epi-, and cis-inositol. As these names suggest, 1L- and 1D-chiro-inositol are the only pair of inositol enantiomers, but they are enantiomers of each other and not enantiomers of myo-inositol. It is understood that any exemplary inositol phosphates shown in this disclosure are not limited to the representative stereostructures shown. Therefore, for example, the examples shown in Figure 3, the silo-, muco-, 1D-chiro-, 1L-chiro-, neo-inositol, allo-, epi-, and cis-inositol stereostructures should also encompass their corresponding equivalents. In most of its stable stereostructures, myo-inositol isomers adopt a chair conformation in which the largest number of hydroxyls are moved to the equatorial position, and these are furthest apart from each other. In this stereostructure, the natural myo-isomer has five of its six hydroxyls (first, third, fourth, fifth, and sixth) in the equatorial position, while the second hydroxyl group is... Axial It has the following structure.

[0058] [ka]

[0059] 2.1. Inositol phosphate, analogues and derivatives

[0073] In some embodiments, the R of a compound of general formula I 1、 R 3、 R 5、 R 7、 R9 and R 11 At least one of these independently represents H, -X, -OX, -NHX, -NX2, -SX, -OSO3HX, -OSO3X2 or a compound of formula II, formula III, or formula IV, where each X independently represents H, C 1~30 Alkyl, C 2~30 Represents alkynyl or Cy1, C 1~30Alkyl, C 2~30 Alkenyl and C 2~30 Alkinyl independently produces one or more R 14 In some cases, Cy1 is replaced by one or more R 15 Optionally substituted by; Cy1 represents a 3- to 10-membered carbocyclic or heterocyclic ring, which can be saturated, partially unsaturated, or aromatic, and the heterocyclic ring has 1 to 4 heteroatoms selected from among O, S, and N, and the ring can be bonded to the rest of the molecule via any available C atoms; Cy1 is optionally condensed into 1 to 4 5- or 6-membered rings, which are saturated, partially unsaturated, or aromatic, carbocyclic, or heterocyclic, and the condensed heterocyclic ring may contain 1 or 2 heteroatoms selected from among O, N, and S; each R 13 H and C are independent of each other. 1~30 Alkyl, -NH2, -NHC 1~30 Alkyl or N(C) 1~30 Alkyl)2 represents each C 1~30 Alkyl groups are independently and optionally substituted with one or more halogen, -OH, -CN, and -NO2 groups; each R 14 and R 15 These are independently -OH, C 1~30 Alkoxy, C 1~30 Alkythionyl, C 1~30 Acyloxy, Phosphate, Halogen, Trihalo C 1~30 This represents alkyl and nitrile azide.

[0060]

[0074] In some implicit ways, each X is independently H, C 1~30 Represents alkyl or Cy1, C 1~30 Alkyl is one or more R 14 In some cases, Cy1 is replaced by one or more R 15 Substituted by in some cases; each R 14 and R 15 These are independently -OH, C 1~30 Alkoxy, C 1~30 Alkylthionyl, C 1~30Acyloxy, Phosphate, Halogen, Trihalo C 1-30 Represents alkyl, nitrile, or azide. In some embodiments, each X is H, C 1~30 It represents an alkyl group or Cy1. 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 H and C are independent of each other. 1~30 Alkyl, -NH2, -NHC 1~30 Alkyl or -N(C) 1~30 Alkyl)2 represents each C 1~30 Alkyl groups are independently and optionally substituted with one or more halogen, -OH, -CN, and -NO2 groups; R2, R4, R6, R8, R 10 and R 12 These independently represent H.

[0062]

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

[0063]

[0077] In additional embodiments, R1, R3, R5, R7, R9 and R 11 At least one of them represents a compound of formula II, formula III, or formula IV, and each R 13 H or C 1~30 Represents alkyl. In another embodiment, R1, R3, R5, R7, R9 and R 11 At least one of them represents a compound of formula II, formula III, or formula IV, and each R 13This represents H.

[0064]

[0078] In detailed 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 IP1, IP2, IP3, IP4, IP6, and combinations of IP6. In some embodiments, IP6 can be dephosphorylated in vivo to form other inositol phosphates (IP5, IP4, IP3, IP2, IP1). Inositol is thought to mean any isomer of the molecule, e.g., myoinositol.

[0065]

[0079] In some embodiments, the compound for use in the present invention is of formula I [wherein,

[0080] R7 is OSO3 - And R9, R5, R3, R1 and R 11 OPO3 2- OPSO2 2- Or OSO3 - Selected independently from;

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

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

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

[0084] R3, R1 and R 11 OPO3 2-, wherein R9, R7 and R5 are OSO3 - ;

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

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

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

[0088] R7 and R5 are OSO3 - , wherein R9, R3, R1, and R 11 is OPO3 2- [which is the case].

[0066]

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

[0067]

[0090] In some aspects, the compound administered at a dose according to the methods disclosed herein is a prodrug that produces the inositol phosphate of the present invention after hydrolysis or other intracellular or extracellular processing.

[0068]

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

[0092] All compounds of formula I contain a group having a COP or COS bond, which confers an affinity to calcium-containing crystals and a sufficiently unstable bond that is hydrolyzable in vivo, thereby preventing irreversible bonding to calcium-containing crystals, such as hydroxyapatite (HAP) in bone. If such irreversible bonding had occurred, it would have adversely affected bone remodeling, as is the case with bisphosphonates when administered long-term because they contain a PCP bond that cannot be hydrolyzed by the body. In other extreme cases, such as phosphorylated compounds that do not contain the COP bond, such as pyrophosphate, their COP bond means that they are too easily hydrolyzed in the intestinal tract, and therefore only parenteral administration is feasible. The compounds of the present invention having COP bonds, COS bonds, and combinations thereof represent a suitable middle ground due to their efficacy and the fact that the body exhibits a mechanism for eliminating the compounds and thus reducing the risk of side effects (for example, compounds with a PCP bond may exhibit a half-life of several months in vivo, thereby affecting bone remodeling, for example).

[0069]

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

[0070]

[0094] The "amine functional group" or "amine group" is an NR'R'' functional group, where R' and R'' are independently selected from hydrogen and C1-C5 alkyl groups. In some embodiments, R' and R'' are selected from hydrogen and C1-C3 alkyl groups. The "hydroxyl functional group" or "hydroxyl 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 - The formula is as follows: "Carboxylic acid amide" is CONR'R'', where R' and R'' independently have the meanings shown above. "Sulfonic acid" is SO3H. "Sulfonic acid amide" is SO2NR'R'', where R' and R'' independently have the meanings shown above.

[0072]

[0096] In the context of this invention, "C1-C3 alkyl" means a saturated linear or branched hydrocarbon having one, two, or three carbon atoms, where one carbon-carbon bond may be unsaturated and one CH2 moiety can be exchanged with oxygen (ether crosslinking). Examples of C1-C3 alkyls that are not limited to this include methyl, ethyl, propyl, propa-2-enyl, and propa-2-inyl.

[0073]

[0097] In the context of the present invention, "C1~C5 alkyl" means a saturated linear or branched hydrocarbon having 1, 2, 3, 4 or 5 carbon atoms, wherein one or two carbon-carbon bonds may be unsaturated, and one CH2 moiety can be replaced with oxygen (ether bridge). Non-limiting examples of C1~C5 alkyl include the examples obtained above for C1~C3 alkyl, further 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. In the context of the present invention, "C3~C 10 alkyl" means a saturated linear or branched hydrocarbon having 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, wherein 1, 2 or 3 carbon-carbon bonds may be unsaturated, and one CH2 moiety can be replaced with oxygen (ether bridge).

[0074]

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

[0075]

[0099] The term "C 2~30 alkenyl" means a linear or branched alkyl chain containing 2 to 30 carbon atoms, which also contains one or more double bonds. Examples include inter alia ethenyl, 1-propenyl, 2-propenyl, isopropenyl, 1-butenyl, 2-butenyl, 3-butenyl and 1,3-butadienyl.

[0076]

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

[0077]

[0101] A "Cy1 group" refers to a 3- to 10-membered carbocyclic or heterocyclic ring that can be saturated, partially unsaturated, or aromatic, and is bonded to the rest of the molecule by any available carbon atoms. If heterocyclic, Cy1 contains 1 to 4 heteroatoms selected from among N, O, and S. Furthermore, Cy1 can optionally be condensed carbocyclic or heterocyclic rings of up to 4, 5-, or 6 members, which can be saturated, partially unsaturated, or aromatic. If the fused ring is heterocyclic, the ring contains 1 or 2 heteroatoms selected from among N, O, and S. Examples of Cy1 include, in particular, 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, pyridadinyl, benzimidazolyl, benzofuranyl, isobenzofuranyl, indolyl, isoindolyl, benzothiophenyl, benzothiazolyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, azetidinyl, and aziridinyl.

[0078]

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

[0079]

[0103] "C" as a base or part of a base 1~30"Alkylthionyl group" refers to -SOC 1~30 It means 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 base or part of a base 1~30 The term "acyloxy group" refers to -COC 1~30 It means alkyl group, C 1~30 The alkyl portion has the same meaning as above. Examples include acetyl, ethanol, propanoyl, and 2,2-diisopropylpentanoyl.

[0081]

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

[0106] "Trihalo C 1~30 An alkyl group, as defined above, is a C group consisting of three halogen groups. 1~30 This refers to a group resulting from the substitution of three hydrogen atoms in an alkyl group. Examples include, among others, trifluoromethyl, tribromomethyl, trichloromethyl, triiodomethyl, trifluoroethyl, tribromoethyl, trichloroethyl, triiodoethyl, tribromopropyl, trichloropropyl, and triiodopropyl.

[0082]

[0107] "-NHC 1~30 "Alkyl alkyl group" is defined as C 1~30 This refers to a group resulting from the substitution of a single hydrogen atom in a -NH2 group by an alkyl group. Examples include, among others, methylamine, ethylamine, propylamine, butylamine, and pentylamine.

[0083]

[0108] "-N(C 1~30 "Alkyl alkyl group" is defined as C 1~30This refers to a group resulting from the substitution of two hydrogen atoms in a -NH2 group by an alkyl group. Examples include, among others, dimethylamine, diethylamine, diisopropylamine, dibutylamine, and diisobutylamine.

[0084]

[0109] The phrase "possibly substituted by one or more" indicates the possibility that a group can be substituted by one or more (e.g., one, two, three, or four) substituents. In some embodiments, a group can be substituted by one, two, or three substituents, and further by one or two substituents, if the group has a sufficient number of available substituted positions. If present, the substituents may be the same or different, and may be at any available position.

[0085]

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

[0086]

[0111] In some embodiments, inositol phosphate, inositol phosphate analogs, and their derivatives are formulas (VIII), (IX), or (X)

[0087] [ka]

[0088] It contains the compound.

[0112] [In the formula, each X is independently OPO3 2- OPSO2 2- , or OSO3 - Selected from; Z is an alkyl chain containing 1 to 3 carbon and / or heteroatoms, and possibly an X group, where X is also OPO3 2- OPSO2 2- , or OSO3 - It was ultimately selected from; R1 (See Section 2.2 below) a heterogeneous structural part, which may be used depending on the circumstances. In some embodiments, a molecule contains two or more heterogeneous structural parts, which may be the same or different.

[0089]

[0113] In some embodiments, Z is CH2, CHX, CHR, as used in equation (VIII). 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 - Selected from.

[0090]

[0114] In some embodiments, Z is (CHX) as used in equation (VIII) p CHX (CHX) q p and q each have values ​​from 0 to 2, independently of each other, except that (p+q) has values ​​of 0, 1, or 2; one, two, or three X can be parts of a heterogeneous structure (e.g., PEG), and the remaining X is OPO3 2- OPSO2 2- , and OSO3 - It is selected independently from. In some embodiments, X of Z is all OPO3 2- This is not necessarily true. In some embodiments, all of Z's X are OSO3. - That is not necessarily true.

[0091]

[0115] In some embodiments, one, two, or three X in a compound of formula (VIII), formula (IX), or formula (X) may be heterostructured parts, and the remaining X may be OPO3 2- OPSO2 2- , or OSO3 - It can be selected independently of the others.

[0092]

[0116] Formula (VII) above describes a 5-membered, 6-membered, or 7-membered alkyl ring, and one or more heterogeneous structural parts, as may be used, are bonded to one of the carbon atoms forming the ring.

[0093]

[0117] In some embodiments, for example, inositol phosphate, 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 OSO3 - X 1 , X 3 , X 4 , X 5 and X 6 OPO3 2- OPSO2 2- Or OSO3 - Selected independently from;

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

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

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

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

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

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

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

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

[0096]

[0128] In some embodiments, the inositol phosphates of the present invention or metabolites thereof can be detected and / or quantified using the method disclosed in US9612250. See also US8377909, US8778912, and US20070066574.

[0097]

[0129] The compounds disclosed herein can exist in any form generally used in pharmaceutical technology. Specific embodiments include, but are not limited to, sodium salts, magnesium salts, potassium salts, ammonium salts, free acids, or mixtures of the foregoing. Other pharmaceutically acceptable salts are known to those skilled in the art and can be readily obtained. In specific embodiments, the compound for use as defined in the first aspect of the present invention is a sodium salt, for example, inositol hexaphosphate hexasodium salt.

[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 myo-inositol. A specific example of a compound for use in the present invention is myo-inositol hexaphosphate hexasodium salt. Sodium salts provide several advantages with respect to the preparation of the resulting IP6 formulation and the level of impurities in the resulting IP6 formulation.

[0099] 2.2. Parts with different structures

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

[0100] [ka]

[0101] Selected from,

[0132] n is an integer in the range of 2 to 200, and R 13 This is selected from H, methyl, or ethyl.

[0102]

[0133] In some embodiments, the compounds for use in the present invention, for example, the inositol phosphate derivatives of the present invention, may contain one or more groups selected from the groups of formulas V, VI, and VII. These groups are heterostructure parts that impart advantageous properties with respect to the corresponding molecule lacking such one or more heterostructure parts. Examples of such advantageous properties that can be imparted to inositol phosphate or inositol phosphate analogs by heterostructure parts or combinations thereof include, but are not limited to, (a) increased solubility, (b) decreased degradation or metabolic rate, (c) increased plasma half-life, (d) decreased hepatic metabolic rate, (e) decreased clearance rate, (f) decreased toxicity, (g) decreased irritability, and (h) decreased side effects. These advantageous properties can be evaluated or quantified using methods known in the art without excessive experimentation.

[0103]

[0134] In some embodiments, the heterosynthetic portion is, for example, polyethylene glycol (PEG) or polyglycerin (PG). Thus, in some embodiments, the compound for use in the present invention is any of the compounds defined in the embodiments disclosed above, including the heterosynthetic portion, i.e., one of the groups of formula I is selected from the groups of formulas V, VI, and VII. In some embodiments, the heterosynthetic portion includes polyethylene glycol (PEG). In some embodiments, the heterosynthetic portion 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. 11At least one of them is a group of formula V. Alternatively, the heterosynthetic portion contains polyglycerin. In some embodiments, the heterosynthetic portion is composed of polyglycerin, i.e., R1, R3, R5, R7, R9 and R of the compound of formula I according to the first embodiment of the present invention. 11 At least one of these is selected from the groups of formula VI or VII. In other embodiments, the compound of formula I according to the first embodiment of the present invention contains one, two, or three groups selected from the groups of formula VI or VII, e.g., two types of PEG (groups of formula V), three types of PEG, three types of polyglycerin (groups of formula VI), three types of PG, or any combination thereof, e.g., one type of PEG and one type of PG, or two types of PEG and one type of polyglycerin. 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 embodiment of the present invention contains two groups selected from the groups of formula VI or VII, e.g., two types of PEG (groups of formula V) or three types of polyglycerin (groups of formula VI) or one type of PEG and one type of polyglycerin, and the residues are all the groups of formula II. In some embodiments, R3 and R7 of the compound of formula I are selected from the groups of formulas V, VI, and VII. In some embodiments, R3 and R7 of the compound of formula I are the groups of formula V, and R1, R5, R9, and R of the compound of formula I. 11 This is the basis of equation II.

[0104]

[0135] The base of formulas V, VI, and VII is R 13 =H, which is methyl or ethyl, and n is an integer from 2 to 200. In some embodiments, R 13= H. In a detailed embodiment, 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 2-10, 10-20, 20-30, 30-40, 40-50, 50-60, 60-70, 70-80, 80-90, 90-100, 100-110, 110-120, 120-130, 130-140, 140-150, 150-160, 160-170, 170-180, 180-190, or 190-200.

[0106]

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

[0138] In some embodiments, PEG is branched PEG. Branched PEG has 3 to 10 PEG chains emanating from a central nuclei.

[0107]

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

[0108]

[0140] In some embodiments, PEG is Comb PEG. Comb PEG has multiple PEG chains, which are usually implanted onto a polymer backbone.

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

[0109]

[0142] In some embodiments, PEG is PEG 100 PEG 200 PEG 300 PEG 400 PEG 500 PEG 600 PEG 700 PEG 800 PEG 900 PEG 1000 PEG 1100 PEG 1200PEG 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 detailed embodiment, PEG is PEG 400 In another detailed aspect, PEG is PEG 2000 That is the case.

[0110]

[0143] In other detailed embodiments, R3 and / or R7 of the compound of formula I are groups of formula V, and 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 the groups of formula V, and R 13 H is an integer between 9 and 45, and R1, R5, R9 and R 11 R is all the groups of formula II. In other detailed embodiments, the compound of formula I is R3 and R7 = the groups of formula V [wherein R 13 H is an integer between 9 and 45, and R1, R5, R9 and R 11 It is a sodium salt having all the groups of formula II.

[0111]

[0144] In some other embodiments, the heterogeneous structural part is represented by the formula ((R3-O-(CH2-CHOH-CH2O)n Polyglycerin (PG) as described by formula (R3) [wherein 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 to these embodiments, n has a value from 9 to 45. In some embodiments, the heterogeneous structural part is formula (R3) 3 -O-(CH2-CHOR 5 -CH2-O) n -) [where R 5 [is hydrogen] Branched polyglycerin or formula (R 3 -O-(CH2-CHOH-CH2-O) n -) [where R 3 It is a linear glycerol described by [where is hydrogen, methyl, or ethyl]. In some embodiments, the heterogeneous structural part is a linear glycerol described by formula (R 3 -O-(CH2-CHOR 5 -CH2-O) n -) [where R 5 [is hydrogen] Superbranched polyglycerin or formula (R 3 -O-(CH2-CHOR 6 -CH2-O) n -) [where R 6 [is hydrogen] described by the glycerol chain or formula (R 3 -O-(CH2-CHOR 7 -CH2-O) n -) [where R 7 [is hydrogen] described by the glycerol chain or formula (R 3 -O-(CH2-CHOH-CH2-O) n -) [where R 3 This refers to linear glycerols described by [the group being hydrogen, methyl, or ethyl]. Superbranched glycerols and methods for their synthesis are known in the art. See Oudshorn M et al., Biomaterials 2006; vol. 27: 5471-5479, Wilms D et al., Acc Chem Res 2010; vol. 43: 129-141 and the references cited therein.

[0112]

[0145] In some embodiments, the molar mass of PG is 100 g / mol to 3000 g / mol, particularly 100 g / mol to 2500 g / mol, and more specifically, approximately 100 g / mol to 2000 g / mol. In some embodiments, the molar mass of PG is 200 g / mol to 3000 g / mol, particularly 300 g / mol to 2500 g / mol, and more specifically, approximately 400 g / mol to 2000 g / mol.

[0113]

[0146] In some aspects, PG is PG 100 PG 200 PG 300 PG 400 PG 500 PG 600 PG 700 PG 800 PG 900 PG 1000 PG 1100 PG 1200 PG 1300 PG 1400 PG 1500 PG 1600 PG 1700 PG 1800 PG 1900 PG 2000 PG 2100 PG 2200 PG 2300 PG 2400 PG 2500 PG 1600 PG 1700 PG 1800 PG 1900 PG 2000 PG 2100 PG 2200 PG 2300 PG 2400 PG 2500 PG 2600 PG 2700 PG 2800 PG 2900 , or PG 3000 In one detailed aspect, PG is PG 400 In another detailed aspect, PG is PG 2000 That is the case.

[0114]

[0147] In other detailed embodiments, R3 and / or R7 of the compound of formula I are groups of formula VI, and 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 the groups of formula VI, and R 13 H is an integer between 9 and 45, and R1, R5, R9 and R 11 R is all the groups of formula II. In other detailed embodiments, the compound of formula I is R3 and R7 = the groups of formula VI [wherein R is the group of formula VI]. 13 H is an integer between 9 and 45, and R1, R5, R9 and R 11 It is a sodium salt having all the groups of formula II.

[0115] 3. Pharmaceutical Compositions

[0148] In other embodiments, the present invention also means a pharmaceutical composition comprising a compound defined in any of the embodiments disclosed above. In some embodiments, the pharmaceutical composition comprises a compound defined in any of the embodiments disclosed above, together with one or more pharmaceutically acceptable excipients or carriers. These pharmaceutical compositions are for use in increasing tissue perfusion and / or oxygen supply in subjects requiring increased tissue perfusion and / or oxygen supply. In some embodiments, these pharmaceutical compositions are for use in the treatment or prevention of ischemia and / or ischemia-related diseases or conditions. In some embodiments, the pharmaceutical compositions of the present invention are used for the treatment or prevention of PAD or CLI.

[0116]

[0149] As used herein, the term “excipient” means a substance that helps the absorption of elements of a pharmaceutical composition, stabilizes such elements, activates a composition, or assists in the preparation of a composition. Therefore, examples of excipients used in parenteral formulations include, but are not limited to, antimicrobial agents (e.g., benzalkonium chloride, metacresol, thimerosal), co-solvents (e.g., ethanol), buffers, and pH adjusters (e.g., carbonic acid, citric acid, phosphoric acid 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 constituent components contained therein to a determined volume or weight. A pharmaceutically acceptable vehicle is an inert substance or a substance having an action similar to that of any of the components comprising the pharmaceutical composition of the present invention. The role of said vehicle is to enable incorporation of other components, to enable better dosing and administration, or to provide consistency and shape to the composition.

[0118]

[0151] The pharmaceutical composition can account for approximately 1% to approximately 95% of the compound as defined in any of the embodiments disclosed above. In some embodiments, the pharmaceutical composition of the present invention can account for, for example, approximately 20% 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% of the compound as defined in any of the embodiments disclosed above.

[0119]

[0152] In some embodiments, the concentration of the inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate, or an analog or derivative thereof, or a combination thereof) in each dose of the pharmaceutical composition is from about 12.5 mM to about 135 mM. In some versions of this embodiment, the concentration of the inositol phosphate of the present 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] Pharmaceutical compositions suitable for parenteral administration include compounds as defined in any of the embodiments disclosed above, mixed with a pharmaceutically acceptable carrier (e.g., sterile water or sterile isotonic saline). Such compositions may be prepared, packaged or sold in forms suitable for rapid or continuous administration. Injectable compositions may be prepared, packaged or sold in unit dosage forms, for example, in the form of ampoules or in the form of multi-dose containers containing preservatives. Compositions for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pasteurized formulations, and implantable sustained-release or biodegradable formulations. Such compositions may further include, but are not limited to, one or more additional components, including suspending agents, stabilizers, or dispersants.

[0121]

[0154] In some embodiments, in a formulation for parenteral administration, the active agent (e.g., a compound as defined in any of the embodiments disclosed above) is provided in a dry (i.e., powder or granular) form for reconstitution with a suitable vehicle (e.g., sterile, pyrogenic water) before parenteral administration of the redissolved composition.

[0122]

[0155] Pharmaceutical compositions may be prepared, packaged, or sold in the form of sterile, injectable aqueous or oily suspensions or solutions. These suspensions or solutions may be formulated according to the known art and may contain, in addition to the active agent (e.g., a compound as defined in any of the embodiments disclosed above), additional components, such as dispersants, wetting agents, or suspending agents as described herein. Such sterile injectable formulations may 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 solutions, and fixative oils, such as synthetic mono- or di-glycerides.

[0123]

[0156] Other useful parentally administered formulations include those containing an effective agent (e.g., a compound as defined in any of the embodiments disclosed above) in microcrystalline form, as a liposomal formulation, or as a component of a biodegradable polymer system.

[0124]

[0157] Compositions for sustained release or transplantation may contain pharmaceutically acceptable polymers or hydrophobic substances, such as emulsions, ion exchange resins, slightly soluble polymers, or slightly soluble salts.

[0125]

[0158] Controlled-release or sustained-release formulations of the pharmaceutical compositions of the present invention can be prepared using prior art. In some cases, the dosage form to be used may provide one or more effective agents as slow-release or controlled-release in various proportions, for example, using hydroxypropyl methylcellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, or microspheres or combinations thereof, to provide a desired release profile. Suitable controlled-release formulations known in the art, including those described herein, can be readily selected for use with the pharmaceutical compositions of the present invention. Accordingly, single unit dosage forms suitable for parenteral or topical administration, such as injectable solutions, gels, creams, and ointments, adapted for controlled-release, are encompassed by the present invention.

[0126]

[0159] Most controlled-release pharmaceutical products share the common goal of improving therapy compared to what is achieved by their uncontrolled counterparts. Ideally, the use of optimally designed controlled-release formulations in medical treatments is characterized by the minimum amount of therapeutic agent used to cure or manage a condition in the minimum amount of time. Advantages of controlled-release formulations include extended therapeutic activity, reduced dosage, and improved patient compliance. Furthermore, controlled-release formulations can be used to influence the onset time of action or other characteristics, such as the blood level of the therapeutic agent, and therefore the occurrence of side effects.

[0127]

[0160] Most controlled-release formulations are designed to release a certain amount of the therapeutic agent first to rapidly produce the desired therapeutic effect, and then release the other amount of the therapeutic agent gradually and continuously to maintain this level of therapeutic effect over an extended period. To maintain this steady level of the therapeutic agent in the body, the therapeutic agent must be released from the dosage form at a rate that replaces the amount of the therapeutic agent that is metabolized or excreted from the body.

[0128]

[0161] The controlled release of an effective drug can be stimulated by various inducers, such as pH, temperature, enzymes, water, or other physiological conditions or compounds. In the context of this invention, the term “controlled-release component” 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 effective drug.

[0129]

[0162] In some embodiments, the formulations of the present invention may, but not limited to, be short-term, rapid-release, and controlled-release formulations, such as sustained-release, delayed-release, and pulsatile-release formulations.

[0130]

[0163] The term sustained-release is used in its conventional sense to mean a therapeutic formulation (e.g., a compound as defined in any of the embodiments disclosed above) that is provided for the gradual release of a therapeutically effective drug over an extended period and, though not necessarily, can result in substantially steady blood levels of the therapeutic agent over that extended period. The period may be one month or longer, and the longer release should be the same amount of the drug administered in the form of rapid dosing.

[0131]

[0164] In the case of sustained release, the compound can be formulated with a suitable polymer or hydrophobic substance that provides sustained-release properties to the compound. Therefore, the compound for use in the method of the present invention can be administered in the form of fine particles, for example, in the form of wafers or disks by injection or implantation. In some embodiments, the compound of the present invention is administered to a patient alone or in combination with another pharmaceutical product using a sustained-release formulation.

[0132]

[0165] The term "delayed release" is used herein in its conventional sense to mean a therapeutic formulation that provides initial release of the therapeutic agent after a delay of some time following administration of the therapeutic agent. The delay may range from about 10 minutes to about 12 hours. The term "pulsatile release" is used herein in its conventional sense to mean a therapeutic formulation that releases the therapeutic agent in such a manner that it results in a pulsed 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 release of the therapeutic agent immediately after administration.

[0133]

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

[0134]

[0167] The pharmaceutical product according to the present invention is manufactured by methods known in the art, particularly by conventional mixing, coating, granulation, dissolution, or freeze-drying.

[0168] The present invention also provides compounds, combinations of compounds, or pharmaceutical formulations as defined in any of the above embodiments of the present invention, either in the broadest sense or as specified in any of the embodiments described above for use as pharmaceuticals.

[0135] 4. Method and route of administration

[0169] In some embodiments, the compounds, pharmaceutical compositions, or combination formulations defined in any of the embodiments disclosed above are administered together, simultaneously, or sequentially with another therapeutic agent. In some versions of these embodiments, the additional therapeutic agent includes cilostazol, pentoxifylline, or a combination thereof.

[0136]

[0170] In some embodiments, an effective dose of a compound, pharmaceutical composition, or combination formulation defined in any of the above embodiments is provided. The compound, pharmaceutical composition, or combination formulation can be administered parenterally, for example, intravenously, intraperitoneally, intramuscularly, intra-arterially, intradermally, subarachnoidally, epidurally, or spinally or subcutaneously. Parenteral administration may be by bolus administration or by intravenous infusion.

[0137]

[0171] In a detailed aspect of the present invention, myo-inositol hexaphosphate (or a formulation containing myo-inositol hexaphosphate, e.g., SNF472, etc.) is administered by intravenous infusion. In another detailed aspect of the present invention, myo-inositol hexaphosphate is administered subcutaneously. In another aspect, an inositol derivative or a myo-inositol hexaphosphate derivative, e.g., R3 and R7 = the group of formula V [wherein R3, R7 = 13 H is an integer between 2 and 200, and R1, R5, R9 and R 11 Compounds of formula I having all the groups of formula II (or their sodium salts) are administered by intravenous infusion. In another embodiment, inositol derivatives or myo-inositol hexaphosphate derivatives, for example, R3 and R7 = the groups of formula V [wherein R is R 13 H is an integer between 2 and 200, and R1, R5, R9 and R 11 Compounds of formula I having all the groups (or their sodium salts) of formula II are administered subcutaneously.

[0138]

[0172] Alternatively, the compound, pharmaceutical composition, or combination formulation may be administered as a component of a hemodialysis, hemofiltration, or peritoneal dialysis solution or system.

[0173] In certain cases of patients treated by dialysis, a very suitable method of administration is to administer the inositol phosphate of the present invention by a dialysis machine (before or after filtration) (e.g., non-bolus administration) instead of directly injecting the inositol phosphate of the present invention into the patient intravenously. Thus, when the patient is circulated through a dialysis circuit while remaining in a dialysis state, and when the blood containing the inositol phosphate of the present invention is returned to the body, the blood can be treated with the inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate).

[0139]

[0174] Accordingly, in some embodiments, the compound, pharmaceutical composition, or combination formulation defined in any of the embodiments disclosed above is administered to the patient during hemodialysis. In some embodiments, the compound, pharmaceutical composition, or combination formulation defined in any of the embodiments disclosed above 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 patient's unfiltered blood in the dialysis circuit). In some embodiments, the compound is inositol hexaphosphate, in particular myo-inositol hexaphosphate sodium salt or derivatives thereof, i.e., R3 and R7 = the group of formula V [wherein R3, R7 = 13 H is an integer between 2 and 200, and R1, R5, R9 and R 11 It is a compound of formula I having all the groups (or their sodium salts) of formula II.

[0140]

[0175] In the case of dialysis patients, administration of the inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate) by a dialysis machine equilibrates the blood with dialysate before returning it to the body; therefore, the inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate) can capture ionized calcium, but this fact is compensated for by the fact that the blood passes through a dialysis filter, thereby removing the aforementioned side effects and significantly improving the safety profile. Furthermore, by administering the inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate) in combination with hemodialysis, particularly when administered to unfiltered blood extracted from the patient during hemodialysis, it becomes possible to reduce the dose of the compound and minimize harmful side effects, which is consequently advantageous in terms of reduced toxicity.

[0141]

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

[0142]

[0177] In general, the effective dose of the inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate), administered according to the methods disclosed herein, depends, 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 dose of the inositol phosphate of the present invention for subjects of several species (e.g., humans) can be calculated based on experimental data available for different or reference species (e.g., rats).

[0143]

[0178] Therefore, for example, a dose of inositol myo-hexaphosphate administered as part of a regimen including administration to rat subjects at a dose of 20 mg / kg should be equivalent to an administration of the same effective drug to human subjects 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 rat subjects should be equivalent to an administration of inositol myo-hexaphosphate at a dose of 8.4 mg / kg to a human subject as defined above. The dosage may be adjusted based on various physiological and psychological factors, including the subject's age, species, body weight, body surface, renal clearance, sex, pathological condition, route of administration, concomitant administration of one or more other drugs, and methods known in the art (Pan S. et al., Patient Prefer Adherence 2016; Vol. 10: 549-560; Pai M, Pharmacotherapy 2012; Vol. 32: 856-868; Hacker M. et al., "Pharmacology: Principles 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 (body weight) of the subject.

[0144]

[0179] In some embodiments, the dose of inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention includes inositol phosphate, inositol phosphate analogs, inositol phosphate derivatives, or combinations thereof according to the present invention, ranging from about 0.001 mg / kg to about 60 mg / kg. In some further embodiments, the dose of inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention is about 0.001 mg / kg to about 20.0 mg / kg, or about 20.0 mg / kg to about 40.0 mg / kg, or about 40.0 mg / kg to about 60.0 mg / kg.

[0145]

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

[0146]

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

[0147]

[0182] In some embodiments, the dose of inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention is approximately 0.001 mg / kg to approximately 0.25 mg / kg, approximately 0.25 mg / kg to approximately 0.5 mg / kg, approximately 0.5 mg / kg to approximately 0.75 mg / kg, approximately 0.75 mg / kg to approximately 1.0 mg / kg, approximately 1.0 mg / kg to approximately 2.50 mg / kg, and approximately 2.50 mg / kg. kg ~ approximately 5.0 mg / kg, approximately 5.0 mg / kg ~ approximately 7.5 mg / kg, approximately 7.5 mg / kg ~ approximately 10.0 mg / kg, approximately 10.0 mg / kg ~ approximately 12.5 mg / kg, approximately 12.5 mg / kg ~ approximately 15.0 mg / kg, approximately 15.0 mg / kg ~ approximately 17.5 mg / kg, approximately 17.5 mg / kg ~ approximately 20.0 mg / kg, approximately 20.0 mg / kg ~ approximately 22.5 mg / kg, approximately 22 0.5 mg / kg to approximately 25.0 mg / kg, approximately 25.0 mg / kg to approximately 27.5 mg / kg, approximately 27.5 mg / kg to approximately 30.0 mg / kg, approximately 30.0 mg / kg to approximately 32.5 mg / kg, approximately 32.5 mg / kg to approximately 35.0 mg / kg, approximately 35.0 mg / kg to approximately 37.5 mg / kg, approximately 37.5 mg / kg to approximately 40.0 mg / kg, approximately 40.0 mg / kg to approximately 42. The dosage ranges are 5 mg / kg, approximately 42.5 mg / kg to 45.0 mg / kg, approximately 45.0 mg / kg to 47.5 mg / kg, approximately 47.5 mg / kg to 50.0 mg / kg, approximately 50.0 mg / kg to 52.5 mg / kg, approximately 52.5 mg / kg to 55.0 mg / kg, approximately 55.0 mg / kg to 57.5 mg / kg, or approximately 57.5 mg / kg to 60.0 mg / kg.

[0148]

[0183] In some embodiments, the dose of inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention is approximately 0.25 mg / kg to approximately 60.0 mg / kg, approximately 0.5 mg / kg to approximately 60.0 mg / kg, approximately 0.75 mg / kg to approximately 60.0 mg / kg, approximately 1.0 mg / kg to approximately 60.0 mg / kg, approximately 2.50 mg / kg to approximately 60.0 mg / kg, and approximately 5 0.0 mg / kg to approximately 60.0 mg / kg, approximately 7.5 mg / kg to approximately 60.0 mg / kg, approximately 10.0 mg / kg to approximately 60.0 mg / kg, approximately 12.5 mg / kg to approximately 60.0 mg / kg, approximately 15.0 mg / kg to approximately 60.0 mg / kg, approximately 17.5 mg / kg to approximately 60.0 mg / kg, approximately 20.0 mg / kg to approximately 60.0 mg / kg, approximately 22.5 mg / kg to approximately 60.0 mg / kg, approximately 25.0 mg / kg to approximately 60.0 mg / kg, approximately 27.5 mg / kg to approximately 60.0 mg / kg, approximately 30.0 mg / kg to approximately 60.0 mg / kg, approximately 32.5 mg / kg to approximately 60.0 mg / kg, approximately 35.0 mg / kg to approximately 60.0 mg / kg, approximately 37.5 mg / kg to approximately 60.0 mg / kg, approximately 40.0 mg / kg to approximately 60.0 mg / kg g, approximately 42.5 mg / kg to approximately 60.0 mg / kg, approximately 45.0 mg / kg to approximately 60.0 mg / kg, approximately 47.5 mg / kg to approximately 60.0 mg / kg, approximately 50.0 mg / kg to approximately 60.0 mg / kg, approximately 52.5 mg / kg to approximately 60.0 mg / kg, approximately 55.0 mg / kg to approximately 60.0 mg / kg, or approximately 57.5 mg / kg to approximately 60.0 mg / kg.

[0149]

[0184] In some embodiments, the dose of inositol phosphate (e.g., myo-inositol hexaphosphate) of the present invention is approximately 0.001 mg / kg to approximately 57.5 mg / kg, approximately 0.001 mg / kg to approximately 55.0 mg / kg, approximately 0.001 mg / kg to approximately 52.5 mg / kg, approximately 0.001 mg / kg to approximately 50.0 mg / kg, approximately 0.001 mg / kg to approximately 47.5 mg / kg, and approximately 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 0.001 mg / kg to about 35.0 mg / kg, about 0.001 mg / kg to about 32.5 mg / kg, about 0.001 mg / kg to about 30.0 mg / kg, about 0.001 mg / 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 The values ​​are approximately g / kg, 0.001 mg / kg to 17.5 mg / kg, 0.001 mg / kg to 15.0 mg / kg, 0.001 mg / kg to 12.5 mg / kg, 0.001 mg / kg to 10.0 mg / kg, 0.001 mg / kg to 7.5 mg / kg, 0.001 mg / kg to 5.0 mg / kg, or 0.001 mg / kg to 2.5 mg / kg.

[0150] 5. Instructions

[0185] The compounds, pharmaceutical compositions, combination formulations, methods of administration, and routes defined in any of the embodiments disclosed above can be used to increase tissue perfusion and / or oxygen supply in subjects requiring such perfusion and / or oxygen supply.

[0151]

[0186] As used herein, the term “ischemic-related disease or condition” means any disease or condition associated with or resulting from an ischemic event or injury. Examples of ischemic-related diseases or conditions 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) diseases or conditions.

[0152]

[0187] In some embodiments, the compounds, pharmaceutical compositions, combination formulations, methods of administration, 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 subjects requiring treatment or prevention thereof.

[0153]

[0188] As used herein, the term “renal failure” means a disease that causes a progressive loss of kidney function, characterized by a simultaneous decrease in the glomerular filtration rate (GFR) or index. Renal failure is also known as renal impairment or kidney disease. Kidney disease can be classified into (i) acute kidney injury (AKI), which is a progressive loss of kidney function, commonly causing oliguria and fluid and electrolyte abnormalities, and (ii) chronic kidney disease (CKD), which is a much slower loss of kidney function over months or years. Depending on the degree of renal function, the 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 ml / min, (d) Stage 4: Severe CKD, GFR = 15-29 ml / min, and (e) Stage 5: End-stage CKD, GFR < 15 ml / min. In Stage 5, dialysis or kidney transplantation is necessary to maintain health. AKI and CKD can occur simultaneously, which is known as an 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 subjects may be acute, chronic, or both. In some embodiments, the subjects are undergoing dialysis (e.g., peritoneal dialysis, hemodialysis). In further embodiments of this embodiment, the subjects are undergoing hemodialysis. In some other embodiments, the subjects are not undergoing dialysis (e.g., subjects with CKD in stages 1-4). In one version of this embodiment, the subjects are administered inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate) 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 subjects may be acute, chronic, or both. In one version of this embodiment, the subjects are undergoing dialysis (e.g., peritoneal dialysis, hemodialysis). In a further embodiment of this version, the subjects are undergoing hemodialysis. In another version of this embodiment, the subjects are not undergoing dialysis (e.g., subjects with CKD in stages 1-4). In one version of this embodiment, the subjects are administered inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate) 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 of administration, and routes of the present invention can be used to improve the walking ability of subjects who require an improvement in walking ability. In some embodiments, the compounds, pharmaceutical compositions, methods of administration, and routes of the present invention can be used to increase maximum walking distance (MWD), maximum walking time (MWT), or both, in subjects who require an increase in these. In some embodiments, the subject suffers from renal disease. The renal disease in the subject may be acute, chronic, or both. In one version of this embodiment, the subject is undergoing dialysis (e.g., peritoneal dialysis, hemodialysis). In a further embodiment 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 inositol phosphate of the present invention (e.g., myo-inositol hexaphosphate) at an effective dose of about 0.001 mg / kg to about 60 mg / kg.

[0157]

[0192] The compounds, pharmaceutical compositions, combination formulations, methods and routes of administration of the present invention are particularly useful for increasing tissue perfusion and / or oxygen supply in the lower extremities, in particular for the treatment and prevention of peripheral artery disease. A further condition that may benefit from the use of inositol phosphate of the present invention is severe limb ischemia. In detailed embodiments, the compounds, pharmaceutical compositions, combination formulations, methods and routes of administration defined in any of the embodiments disclosed above are particularly for use in increasing tissue perfusion and / or oxygen supply in the lower extremities, in particular 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 inositol phosphate of the present 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 pharmaceutically acceptable salt thereof, for use in increasing tissue perfusion and / or oxygen supply in subjects requiring increased tissue perfusion and / or oxygen supply.

[0159] [ka]

[0160]

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

[0161] [ka]

[0162]

[0196] (In the formula, n is an integer in the range of 2 to 200, R 13 H, methyl, ethyl and C3-C 10 Independently selected from the groups (selected from alkyl),

[0197] however:

[0198] R1, R3, R5, R7, R9 and R 11 At least one of these is selected from the bases of equations II, III, and IV,

[0199] R1, R3, R5, R7, R9 and R 11 [The condition is that 0, 1, 2, or 3 of these are selected from the bases of formulas V, VI, and VII.]

[0163]

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

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

[0164]

[0202] Embodiment 4. A compound for use according to any one of Embodiments 1 to 3, for the treatment of a subject undergoing dialysis, preferably hemodialysis.

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

[0165]

[0204] Embodiment 6: R1, R3, R5, R7, R9 and R 11 A compound for use according to any one of Embodiments 1 to 5, wherein at least two, at least three, at least four, at least five, or at least six of the groups are selected from the groups of formulas V, VI, and VII.

[0166]

[0205] Embodiment 7.R 1、 R 3、 R 5、 R 7、 R9 and R 11 A compound for use according to Embodiment 6, wherein at least two, at least three, at least four, at least five, or at least six of the groups are the group 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 - And R1, R3, R5, R9 and R 11 However, OPO3 2- OPSO2 2- Or OSO3 - Selected independently from;

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

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

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

[0213] R3, R1 and R 11 However, OPO3 2- R9, R7, and R5 are OSO3 - and;

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

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

[0216] R7 and R5 are OPO3 2- And R9, R3, R1, and R 11 However, OSO3 - is; or,

[0217] R7 and R5 are OSO3 - And R9, R3, R1, and R 11 However, OPO3 2- The compound for use according to Embodiment 7.

[0169]

[0218] Embodiment 11. A compound for use according to any one of Embodiments 1 to 10, wherein the compound of Formula I has a myo-inositol stereostructure.

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

[0170]

[0220] Embodiment 13. R1, R3, R5, R7, R9 and R 11 Four of these are the bases of Equation II, R1, R3, R5, R7, R9 and R 11 Two of these are selected from the groups of formulas V, VI, and VII, and the compound is for use according to Embodiment 12.

[0171]

[0221] Embodiment 14. R1, R3, R5, R7, R9 and R 11 Four of these are the bases of Equation II, R1, R3, R5, R7, R9 and R 11 Two of these are groups of formula V, a compound for use according to Embodiment 13.

[0172]

[0222] Embodiment 15. (i) R1, R5, R9 and R 11 However, R3 and R7 are selected from the bases of equations V, VI, and VII. (i) R1, R3, R9 and R 11 A compound for use according to any of Embodiments 12 to 14, wherein the group is of Formula II, and R5 and R7 are selected from the groups of Formulas V, VI, and VII.

[0173]

[0223] Embodiment 16. A compound for use according to Embodiment 15, wherein a 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 to 16, wherein the group of formula V, VI, or VI is in the range of 2 to 200, and n is in the range of 2 to 200.

[0174]

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

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

[0175]

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

[0228] Embodiment 21.R 13 A compound for use according to any of embodiments 12 to 20, wherein H is present.

[0176]

[0229] Embodiment 22. R1, R5, R9 and R 11 A compound for use according to Embodiment 21, wherein R3 and R7 are groups 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 pharmaceutically 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 one of Embodiments 23 to 26, in a dried form for reconstitution by a suitable vehicle.

[0179]

[0235] Embodiment 28. A pharmaceutical composition according to any one of Embodiments 23 to 26, wherein the solution is preferably an isotonic saline solution.

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

[0180]

[0237] Embodiment 30. A pharmaceutical composition according to any one of Embodiments 23 to 29, 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, administered to a patient undergoing dialysis.

[0181]

[0239] Embodiment 32. The 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, administered before dialysis.

[0182]

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

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

[0183]

[0243] Embodiment 36. A compound or pharmaceutical composition for use according to any one of Embodiments 31 to 35, administered via a 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 intravenous infusion.

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

[0185]

[0247] Embodiment 40. A compound or pharmaceutical composition for use according to Embodiments 1 to 39, wherein the compound is administered to the subject in a therapeutically effective dose of approximately 0.001 mg / kg to approximately 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 dose of approximately 15 mg / kg to approximately 45 mg / kg.

[0186]

[0249] Embodiment 42.(i)(a) a combination formulation comprising 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 oxygen supply in a subject that requires increased tissue perfusion and / or oxygen supply, comprising the step of 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 that requires treatment or prevention of ischemia, comprising the step of 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 that requires improvement, comprising the step of 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 the maximum walking distance (MWD), maximum walking time (MWT), or both, in a subject that requires an increase in these, comprising the step of 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 artery disease in a subject that requires treatment or prevention of peripheral artery disease, comprising the step of 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. A method according to any one of Embodiments 44 to 48, wherein the combination formulation is 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 cited documents throughout this application are incorporated herein by reference in their entirety.

[0193] General procedure 1. Blood perfusion of the limbs

[0258] Position limb blood perfusion and ischemic state (i.e., blood perfusion including perfusion units, perfusion difference, and perfusion ratio) are evaluated 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 measurement. Perfusion difference and perfusion ratio are calculated by comparing baseline and any indicated intermediate or final measurements for each group. Blood flow is evaluated at C15 minutes after treatment with the effective drug being tested (i.e., 15 minutes after treatment with SNF472, 20 minutes after treatment with IP4-BIS-PEG100, and 3-4 hours after treatment with cilostazol). max Evaluate it around that point.

[0194] 2. Walking ability test

[0259] Walking ability (maximum walking time (MWT) and maximum walking distance (MWD)) is quantified by a forced-increase treadmill running test. A 2-lane rodent treadmill (LE8709TS; PanLab / Harvard Apparatus, Holliston, MA, US) is used. The treadmill is run at a 15% incline, with a speed of 15 m / min (25 cm / sec) for the first 5 minutes, then at 33 cm / sec for the next 5 minutes, and finally at a speed of 40 cm / sec for a maximum of 30 minutes.

[0195]

[0260] Limb function is assessed by the effective drugs being tested (i.e., 15 minutes after treatment with SNF472 and 3-4 hours after treatment with cilostazol). max The subjects will be evaluated based on their performance. Before the test, they will be instructed to exercise on a treadmill according to a pre-test protocol for acclimatization. Subjects who do not follow the protocol will be excluded from the test.

[0196]

[0261] The subjects are made to run for up to 40 minutes or until they become exhausted during the test (i.e., the subjects remain on the shock grid for 5 consecutive seconds). Then, MWD and MWT are calculated for each animal.

[0197] 3. Tissue and blood collection, calcification assay

[0262] The subject is anesthetized by isoflurane inhalation. Blood is obtained by hemorrhage via cardiac puncture. The subject is then euthanized, and its tissues (e.g., right and left femoral arteries, aorta) are collected. The blood and tissues are processed and analyzed to determine their calcium content.

[0198]

[0263] The calcium content in tissue samples is quantified by inductively coupled plasma atomic emission spectroscopy (ICP-OES) using an Optima 7300 DV ICP-OES System spectrometer (PerkinElmer, Inc., Waltham, MA, US) according to the manufacturer's instructions. Myo-inositol-hexaphosphate levels in plasma are quantified by LC-MS / MS chromatography as described in the Art. See WO2013050603.

[0199] Example 1 Prevention of limb ischemia Effects of blood perfusion

[0264] The prophylactic effects of SNF472, IP4-4,6-bisPEG100 sodium salt, and cilostazol on hemoperfusion were tested in a rat model over a 12-day period. Limb ischemia was induced in the subjects from day 1, except for a pseudo-group that did not experience ischemia. Subsequently, subjects with ischemia-induced ischemia were treated with placebo and effective drug combinations from day 1 to day 12, and their effects on the prevention of limb ischemia were evaluated. Observations were made at several points during treatment from day 1 to day 12. All subjects were weighed daily before treatment.

[0200] 1. Induction of limb ischemia

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

[0201]

[0266] Group 1 - Control (Pseudo)

[0267] Group 2a: Placebo - Physiological 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 groups 2a-25, limb ischemia was induced daily from day 1 to day 3 by subcutaneous administration of 120,000 IU / kg of vitamin D3 (cholecalciferol, dufafral D31000; Zoetis Inc., Parsippany, NJ, US) in physiological saline (2 mL / kg). In the first suspected group, physiological saline (2 mL / kg) without vitamin D3 was administered daily from day 1 to day 3.

[0202]

[0273] Subjects in groups 1 and 2a received subcutaneous administration of physiological saline (2 mL / kg) daily from day 1 to day 12. Subjects in group 2b received oral administration of 5% CMC sodium salt in aqueous solution (5 mL / kg) daily from day 1 to day 12.

[0203]

[0274] The administration of vitamin D3, which induced ischemia in the hind limbs of subjects in groups 2a, 2b, and 3-5, was evaluated using laser Doppler perfusion imaging. 2. Effects on ischemia rescue and limb hemoperfusion - Laser Doppler imaging assay

[0275] In subjects in groups 3 and 4, limb hemoperfusion was induced daily from D1 to D12 by subcutaneous administration of 20 mg / kg of SNF472 (Na6IP6; free base: 600 g / mol) and IP4-4,6-bisPEG100 sodium salt (free base 696.27 g / mol), respectively, in physiological saline (2 mL / kg). In subjects in group 5, from D1 to D12, cilostazol (C) was administered daily in aqueous solution of 5% CMC sodium salt (5 mL / kg). 20 H 27 Oral administration of 20 mg / kg of N5O2 (free base 369.46 g / mol; lot number LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) induced limb hemoperfusion.

[0204]

[0276] In all rats, limb ischemia was assessed at D0 (baseline), D6, and D12 by laser Doppler perfusion imaging. Perfusion difference and perfusion ratio were calculated by comparing either the D0 baseline or the D6 and D12 measurements for each group. In particular, perfusion difference and perfusion ratio were calculated by comparing the measurements of Group 1 (control) with (a) Groups 2a and 2b (placebo groups, i.e., physiological saline and 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).

[0205]

[0277] On the days when hemoperfusion tests were performed with the use of effective drugs (i.e., D6 and D12), the following doses were administered:

[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 H 27N5O2): 3-4 hours before measurement

[0281] Under the above scheme, effective drugs are identified by their maximum serum concentration (C) at the time the test measurement is performed. max ) should be the case.

[0206]

[0282] The subjects in Group 1 showed no significant changes in those perfusion parameters.

[0283] Administration of SNF472 and IP4-4,6-bisPEG100 sodium salt attenuated the decrease in hemoperfusion in the limbs of subjects in groups 3–5 (i.e., those 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 hemoperfusion in the hind limbs of treated subjects. See Figures 7, 8, and 9.

[0207] 3. Calcium content and calcification - ICP-OES

[0284] SNF472 suppressed aortic calcification (31±16%) after daily subcutaneous administration of 20 mg / kg compared to placebo, demonstrating its effectiveness against vascular calcification. 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 hemorrhage. They were then autopsied, and their aortas were collected. The tissues were freeze-dried for 24 hours and weighed. The freeze-dried tissues were then immersed in a 1:1 HNO3:HClO4 mixture in a dry bath incubator at 180°C for 2–4 hours. Subsequently, the immersed tissues were diluted to a final volume of 10 mL using ultra-high purity Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US). 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 was collected on day 12. Approximately 8–10 mL of total blood was collected from each subject and divided into blood collection tubes for plasma (K3EDTA, approximately 6 mL of blood) and serum (approximately 2–3 mL of blood) testing. Plasma was stored in one 600 μL aliquot and several other 500 μL aliquots were stored. Serum was divided into two aliquots.

[0210]

[0287] Plasma myo-inositol-hexaphosphate levels in subjects in Group 3 (i.e., C15 minutes after the final SNF472 dose) max The samples (collected at the target location) were quantified by LC-MS / MS chromatography as described in this technical field. See WO2013050603.

[0211]

[0288] All subjects in Group 3 were adequately exposed to the SNF472 product. Plasma levels 15 minutes after the final subcutaneous administration on D12 were 15587 ± ng / mL (24 ± 12 μM). Plasma levels in rats after daily administration of 20 mg / kg were comparable to those found in hemodialysis patients treated intravenously with a dose of 4.2 mg / kg of SNF472.

[0212] Example 2 Prevention of limb ischemia Impact on Maximum Walking Time (MWT) and Distance Walking Time (MDT)

[0289] The prophylactic effects of SNF472 and cilostazol on hemoperfusion, walking ability, and tissue calcification were tested in a rat model over 24 days. Furthermore, the effect of combined SNF472 and cilostazol therapy on hemoperfusion was also examined. Limb ischemia was induced in all subjects from D1 to D3, excluding a pseudo-group that did not experience ischemia. The ischemic subjects were then treated with placebo and an effective drug combination from D1 to D12 to evaluate their effects on (a) preventing limb ischemia and tissue calcification and (b) preventing deterioration of walking ability. Treatment was discontinued in all subjects from D13 to D24. Observations were made at several points during the treatment and post-treatment phases from D1 to D24. All subjects were weighed daily before treatment.

[0213] 1. Induction of limb ischemia

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

[0214]

[0291] Group 1 - Control (Pseudo)

[0292] Group 2a: Placebo - Physiological 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 groups 2a-25, limb ischemia was induced daily from day 1 to day 3 by subcutaneous administration of 120,000 IU / kg of vitamin D3 (cholecalciferol, dufafral D31000; Zoetis Inc., Parsippany, NJ, US) in physiological saline (2 mL / kg). In the first suspected group, physiological saline (2 mL / kg) without vitamin D3 was administered subcutaneously daily from day 1 to day 3.

[0215]

[0298] Subjects in groups 1 and 2a received subcutaneous administration of physiological saline (2 mL / kg) daily from day 1 to day 12. Subjects in group 2b received oral administration of 5% CMC sodium salt in aqueous solution (5 mL / kg) daily from day 1 to day 12.

[0216] 2. Effects on ischemia rescue and hemoperfusion - Laser Doppler imaging assay

[0299] Subjects in groups 1 and 2a were administered 2 mL / kg of physiological saline daily via subcutaneous route from D1 to D12. Subjects in group 2b were administered 5 mL / kg of 5% CMC sodium salt in aqueous solution orally daily from D1 to D12. Furthermore, subjects in group 3 were administered 20 mg / kg of SNF472 (Na6IP6, free base: 600 g / mol) sodium salt (free base 696.27 g / mol) in physiological saline (2 mL / kg) daily via subcutaneous route from D1 to D12. Subjects in group 4 were administered cilostazol (C) in 5% CMC sodium salt in aqueous solution (5 mL / kg) daily from D1 to D12. 20 H 27 N5O2, free base 369.46 g / mol; lot number LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) 20 mg / kg was administered orally. In the subjects of Group 5, (i) cilostazol (C) in 5% CMC sodium salt (5 mL / kg) aqueous solution was administered. 20 H 27(ii) 20 mg / kg of N5O2 (free base 369.46 g / mol; lot number LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) was administered orally, followed by (ii) 20 mg / kg of SNF472 (Na6IP6, free base: 600 g / mol) in physiological saline (2 mL / kg) administered subcutaneously. Administration was carried out daily from D1 to D12.

[0217]

[0300] On the days when hemoperfusion tests were performed with the use of effective drugs (i.e., D6, D12, and D18), the following doses were administered:

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

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

[0303] Group 5 - Cilostazol + SNF472: 15 minutes before measurement for SNF472 and 3-4 hours before measurement for cilostazol.

[0304] Under the above scheme, effective drugs are identified by their maximum plasma concentration (C) at the time the test measurement is performed. max ) should be the case.

[0218]

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

[0219]

[0306] SNF472 alone or in combination with cilostazol attenuated limb ischemia in rats. Treatment with cilostazol alone was ineffective in this model. The effect of SNF472 on hemoperfusion was maintained every 6 days after treatment discontinuation. See Figure 11.

[0220] 3. Effects on walking ability - Treadmill running test

[0307] Maximum walking time (MWT) and maximum walking distance (MWD) were evaluated in groups 1-4 at D0, D5, D10, and D17 using treadmill running tests (8-10 animals per group and at each time point).

[0221]

[0308] The subjects were acclimated to a treadmill for two days prior to the experiment. On the first day, the subjects were exercised for 5 to 10 minutes at treadmill speeds ranging from 15 m / min to 24 m / min in a progressive manner. On the second day, the subjects were first exercised at a speed of 15 m / min for 5 minutes. Next, they were exercised at 19.8 m / min (33 cm / sec) for another 5 minutes. Finally, they were exercised at 24 m / min (40 cm / sec) for up to 30 minutes. Pre-operative walking time and distance were recorded. Animals that did not follow the protocol were excluded from the experiment.

[0222]

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

[0223]

[0310] Under the above scheme, effective drugs are measured at their maximum plasma concentration (C) at the time the running test is performed. max ) should be the case.

[0311] The subjects were made to run for 40 minutes or until exhaustion (i.e., the subjects remained on the shock grid for 5 consecutive seconds). Then, MWD and MWT were calculated for each animal.

[0224]

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

[0225] 4. Calcium content and calcification - ICP-OES

[0313] The subjects were anesthetized, and their blood was collected on day 24. The subjects were euthanized after blood loss. Then, autopsies were performed, and their aortas were collected. The tissues were freeze-dried for 24 hours and weighed. The freeze-dried tissues were then immersed in a 1:1 HNO3:HClO4 mixture in a dry bath incubator at 180°C for 2-4 hours. Subsequently, the immersed tissues were diluted to a final volume of 10 mL using ultra-high purity Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US). The calcium content in the tissue samples was quantified by ICP-OES.

[0226]

[0314] SNF472 suppressed aortic calcification (41±9%) after daily subcutaneous administration at 20 mg / kg compared to placebo, demonstrating its effectiveness against vascular calcification. Cilostazol was not active against calcification at the same dose. See Figure 14.

[0227] Example 3 Treatment of limb ischemia

[0315] The effects of SNF472 and cilostazol on hemoperfusion, walking ability, and tissue calcification were tested in a rat model over a 13-day period after the initiation of limb ischemia. Limb ischemia was induced in all groups from D1 to D3, except for a pseudo-group that did not experience ischemia. From D5 onward, ischemia was induced in the ischemic subjects by administering placebo and effective drug combinations. 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 treatment from D1 to D13. All subjects were weighed daily before treatment.

[0228] 1. Induction of limb ischemia

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

[0229]

[0317] Group 1 - Control (Pseudo)

[0318] Group 2 - D5 Ca baseline

[0319] Group 3a: Placebo - Physiological 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] In groups 2-5, limb ischemia was induced daily from day 1 to day 3 by subcutaneous administration of 120,000 IU / kg of vitamin D3 (cholecalciferol, dufafral D31000; Zoetis Inc., Parsippany, NJ, US) in physiological saline (2 mL / kg). In group 1, vitamin D3-free physiological saline (2 mL / kg) was administered daily from day 1 to day 3.

[0230] 2. Effects on ischemia rescue and hemoperfusion - Laser Doppler imaging assay

[0324] In Group 1, subjects were administered 2 mL / kg of physiological saline daily via subcutaneous route from D1 to D13. In Group 2, subjects were administered 2 mL / kg of physiological saline daily via subcutaneous route from D1 to D5. On D5, four subjects from Group 1 and all subjects from Group 2 were sacrificed to determine their baseline Ca values.

[0231]

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

[0232]

[0326] Among the subjects in Group 5, cilostazol (C) in 5% CMC sodium salt (5 mL / kg) aqueous solution was used. 20 H 27 N5O2, free base 369.46 g / mol; lot number LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) 40 mg / kg was administered orally daily from D5 to D13. A 40 mg / kg dose of cilostazol in rats is equivalent to a therapeutic dose of 8.4 mg / kg in PAD patients.

[0233]

[0327] On the days when hemoperfusion tests were performed with the use of effective drugs (i.e., D5 and D13), the following doses were administered:

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

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

[0330] Under the above scheme, effective drugs are identified by their maximum plasma concentration (C) at the time the test measurement is performed. max ) should be the case.

[0234]

[0331] Limb function and ischemia were evaluated 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 baseline and either D5 or D13 measurements for each group. In particular, perfusion difference and perfusion ratio were calculated by comparing measurements for group 1 (control) with (a) placebo groups 3a and 3b (physiological saline, 5% CMC sodium salt solution), (b) group 4 (SNF472), and (c) group 5 (cilostazol).

[0235]

[0332] VitD3 administration induced a decrease in hindlimb hemoperfusion (measured on D5 immediately prior to therapy administration) in groups 3a, 3b, 4, and 5. Only animals treated with SNF472 showed a significant improvement in limb hemoperfusion on D13 compared to pre-treatment D5. Compared to D5, animals treated with placebo or cilostazol on D13 showed no improvement or reported ischemic rescue. See Figure 16.

[0236] 3. Effects on walking ability - Treadmill running test

[0333] Maximum walking time (MWT) and maximum walking distance (MWD) were evaluated in all groups on D0, and in groups 1, 3a, 3b, 4, and 5 on D6 and D11 using treadmill running tests (8-12 animals per group and at each time point). Participants were acclimatized to the treadmill for two days prior to the test. On the first day, participants were exercised for 5-10 minutes at progressively increasing treadmill speeds ranging from 15 m / min to 24 m / min. On the second day, participants were first exercised at 15 m / min for 5 minutes, followed by another 5 minutes at 19.8 m / min (33 cm / sec), and finally, up to 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 assessed in groups 1, 3a, 3b, and 4 using a treadmill running test 15 minutes after the corresponding daily dose. Limb function in group 5 was assessed 3–4 hours after treatment.

[0238]

[0335] Under the above scheme, effective drugs are identified by their maximum plasma concentration (C) at the time the test measurement is performed. max ) should be the case.

[0336] The subjects were made to run for 40 minutes or until exhaustion (i.e., the subjects remained on the shock grid for 5 consecutive seconds). Then, MWD and MWT were calculated for each animal.

[0239]

[0337] Even when treatment was initiated 5 days after ischemic induction, SNF472 improved walking ability in rats compared to vehicle-based rats (+49% MWD), while cilostazol was ineffective under the same conditions and at therapeutic doses (40 mg / kg / day). See Figure 16.

[0240] 4. Calcium content and calcification - ICP-OES

[0338] The four subjects from Group 1 and all subjects from Group 2 were sacrificed on D5 to determine their baseline Ca values. The remaining subjects from Group 1, as well as all subjects from Groups 3a, 3b, 4, 5, and 6, were sacrificed on D13.

[0241]

[0339] The subjects were euthanized after hemorrhage. Necropsies were then performed, and the right and left femoral arteries were collected. The tissues were freeze-dried for 24 hours and weighed. The freeze-dried tissues were then immersed in a 1:1 HNO3:HClO4 mixture in a dry bath incubator at 180°C for 2–4 hours. Subsequently, the immersed tissues were diluted to a final volume of 10 mL using ultra-high purity Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US). Calcium content in the tissue samples was quantified by ICP-OES.

[0242]

[0340] SNF472 was shown to be effective against vascular calcification on day 13. Compared to placebo, SNF472 suppressed calcification in the femoral artery up to 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 was collected on day 13. Approximately 8-10 mL of total blood was collected from each subject and divided into plasma (K3EDTA, approximately 6 mL of blood) and serum (approximately 2-3 mL of blood) in blood collection tubes for testing. The plasma was stored as one 600 μL aliquot and several other 500 μL aliquots. The serum was divided into two aliquots.

[0244]

[0342] Plasma myo-inositol-hexaphosphate levels (i.e., C15 minutes after the final SNF472 dose) of subjects in Group 4. max The samples (collected at the target location) were quantified by LC-MS / MS chromatography as described in this technical field. See WO2013050603.

[0245]

[0343] All subjects in Group 4 were adequately exposed to the SNF472 product. Plasma levels 15 minutes after the final subcutaneous administration on D13 were 40078 ± 15024 ng / mL (60.7 ± 22.8 μM). Plasma levels in rats after daily administration of 20 mg / kg were comparable to those found in hemodialysis patients treated with SNF472 at a dose of 8.4 mg / kg via intravenous route.

[0246] Example 4 SN472 Drug Interactions

[0344] We analyzed the compatibility of SNF472 with other drugs regularly prescribed to patients with impaired renal function.

[0247]

[0345] Wistar rats (Charles River Labs, Inc., Wilmington, MA, US) were treated as follows: (i) subcutaneous administration of SNF472 (sc), (ii) oral administration of SNF472 + sevelamer, (iii) oral administration of SNF472 (sc) + cinacalcet, (iv) oral administration of SNF472 (sc) + vitamin D, (v) sodium thiosulfate (sc), and ibandronate (sc). No significant difference was observed between administration of SNF472 alone or in combination with any of the other drugs being assayed.

[0248] Example 5 Prevention of limb ischemia The dose-response effect of SNF472 on maximum walking time (MWT) and distance time (MDT).

[0346] The prophylactic effects of several different doses of SNF472 and the maximum permissible dose of cilostazol on hemoperfusion, walking ability, and tissue calcification were tested over 12 days in a rat model. Except for the surrogate group, limb ischemia was induced in the subjects from D1 to D3. The ischemic-induced animals were then treated with placebo and effective drug combinations from D1 to D12 to evaluate their effects on (a) preventing limb ischemia and tissue calcification and (b) preventing deterioration of walking ability. Observations were made at several points during the treatment phases from D1 to D12. All subjects were weighed daily before treatment.

[0249] 1. Induction of limb ischemia

[0347] We used 102 male Sprague Dolly (SD) rats (Envigo Corp., Huntingdon, GB) weighing approximately 250-275g. The rats were fed AO4 diet (Scientific Animal Food & Engineering; Carpe Bio, Amersfoort, NL). The rats were divided into 5 groups of 8-12 animals each, as follows:

[0250]

[0348] Group 1 - Control (Pseudo)

[0349] Group 2a: Placebo - Subcutaneous saline solution

[0350] Group 2b: Placebo - 5% CMC sodium salt solution, orally.

[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) 45 mg / kg, orally

[0357] In groups 2-8, limb ischemia was induced daily from day 1 to day 3 by subcutaneous administration of 120,000 IU / kg of vitamin D3 (cholecalciferol, dufafral D31000; Zoetis Inc., Parsippany, NJ, US) in physiological saline (2 mL / kg). In group 1 (the suspected group), vitamin D3-free physiological saline (2 mL / kg) was administered daily from day 1 to day 3.

[0251]

[0358] Subjects in groups 1 and 2a received subcutaneous administration of physiological saline (2 mL / kg) daily from day 1 to day 12. Subjects in group 2b received oral administration of 5% CMC sodium salt in aqueous solution (5 mL / kg) daily from day 1 to day 12.

[0252] 2. Effects on ischemia rescue and hemoperfusion - Laser Doppler imaging assay

[0359] Subjects in groups 1 and 2a were administered 2 mL / kg of physiological saline daily via the subcutaneous route from D1 to D12. Subjects in group 2b were administered 5 mL / kg of 5% CMC sodium salt in aqueous solution orally daily from D1 to D12. Furthermore, subjects in groups 3, 4, 5, 6, and 7 were administered SNF472 (Na6IP6, free base: 600 g / mol) sodium salt (free base 696.27 g / mol) at doses of 1 mg / kg, 7.5 mg / kg, 15 mg / kg, 30 mg / kg, and 45 mg / kg, respectively. SNF472 was administered daily from D1 to D12 via the subcutaneous route in physiological saline (2 mL / kg). Subjects in group 8 were administered cilostazol (C) in 5% CMC sodium salt in aqueous solution (5 mL / kg). 20 H 27 N5O2, free base 369.46 g / mol; lot number LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) 45 mg / kg was administered orally daily from D1 to D12.

[0253]

[0360] On the days when hemoperfusion tests were performed with the use of effective drugs (i.e., D6 and D12), the following doses were administered:

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

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

[0363] Under the above scheme, effective drugs are identified by their maximum plasma concentration (C) at the time the test measurement is performed. max ) should be the case.

[0254]

[0364] Limb ischemia was assessed in all rats during the treatment period (i.e., D0, D6, and D12) by laser Doppler perfusion imaging. Perfusion difference and perfusion ratio were calculated by comparing baseline and either D6 or D12 measurements for each group. In particular, perfusion difference and perfusion ratio were calculated by comparing measurements from Group 1 (control) with (a) Groups 2a and 2b (placebo groups: physiological saline, 5% CMC sodium salt solution), (b) Groups 3, 4, 5, 6, and 7 (SNF472), and (c) Group 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] Maximum walking time (MWT) and maximum walking distance (MWD) were evaluated in groups 1-8 at D0, D5, and D10 using treadmill running tests (8-12 animals per group and at each time point).

[0256]

[0367] The subjects were acclimated to a treadmill for two days prior to the experiment. On the first day, the subjects were exercised for 5 to 10 minutes at treadmill speeds ranging from 15 m / min to 24 m / min in a progressive manner. On the second day, the subjects were first exercised at a speed of 15 m / min for 5 minutes. Next, they were exercised at 19.8 m / min (33 cm / sec) for another 5 minutes. Finally, they were exercised at 24 m / min (40 cm / sec) for up to 30 minutes. Pre-operative walking time and distance were recorded. Animals that did not follow the protocol were excluded from the experiment.

[0257]

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

[0258]

[0369] Under the above scheme, effective drugs are measured at their maximum plasma concentration (C) at the time the running test is performed. max ) should be the case.

[0370] The subjects were made to run for 40 minutes or until exhaustion (i.e., the subjects remained on the shock grid for 5 consecutive seconds). Then, MWD and MWT were calculated for each animal.

[0259]

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

[0260] 4. Calcium content and calcification - ICP-OES

[0372] The subjects were anesthetized, and their blood was collected on day 12. The subjects were euthanized after blood loss. They were then autopsied, and their hearts and aorta arteries were collected. The tissues were freeze-dried for 24 hours and weighed. The freeze-dried tissues were then immersed in a 1:1 HNO3:HClO4 mixture in a dry bath incubator at 180°C for 2-4 hours. Subsequently, the immersed tissues were diluted to a final volume of 10 mL using ultra-high purity Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US). The calcium content in the tissue samples was quantified by ICP-OES.

[0261]

[0373] SNF472 was shown to be effective in a dose-response manner against cardiac and vascular calcification, such as calcification of the heart and aortic arteries. Cilostazol was not active against calcification at the same dose.

[0262] Example 6 Prevention of limb ischemia in a rat model of uremic disease induced by adenine The effect of SNF472 on limb blood perfusion

[0374] The prophylactic effects of SNF472 and cilostazol doses on hemoperfusion and tissue calcification were tested over 21 days in a rat model of related chronic kidney disease.

[0263]

[0375] Uremia and limb ischemia were induced in animals from D1 to D21, excluding a pseudo-group that did not develop either uremia or ischemia. The ischemic-induced animals were then treated with placebo and effective drug formulations from D1 to D21 to evaluate their effects on (a) preventing limb ischemia and (b) preventing tissue calcification. Several observations were made during the treatment phases from D1 to D21. All subjects were weighed daily before treatment.

[0264] 1. Induction of uremia and limb ischemia

[0376] Sixty-eight male Sprague Dolly (SD) rats (Envigo Corp., Huntingdon, GB), weighing approximately 250-275g, were freely fed a pellet-type high-phosphorus diet (SM R, 10mm solid feed, Ca 1.06%, P 1.03%) (SSNIFF Spezialdiaeten, Soest, DE). The subjects were divided into six groups of 8-12 animals each, as follows:

[0377] Group 1 - Control (Pseudo)

[0378] Group 2a: Placebo - Subcutaneous saline solution, once daily.

[0379] Group 2b: Placebo - Physiological 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 - An Alzet pump containing a total dose of SNF472(Na6IP6) 400 mg / 4 weeks (100 mg / week) was implanted subcutaneously for 4 weeks on D1 before adenine administration.

[0265]

[0383] Group 6 - Cilostazol (C 20 H 27 N5O2) 45 mg / kg / day, orally, once daily.

[0384] In groups 2-6, cardiovascular calcification and ischemia were accelerated and homogenized by inducing adenine at a daily dose (500 mg / kg orally suspended in 1% carboxymethylcellulose) for the first 10 days, followed by α-calcidol at a dose (100 ng / kg orally in olive oil) three times per week from D11 to D19.

[0266]

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

[0267]

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

[0268] 2. Effects on ischemia rescue and hemoperfusion - Laser Doppler imaging assay

[0387] In group 2a, subjects received subcutaneous administration of physiological saline (2 mL / kg) twice daily for 21 days. In group 2b, subjects received subcutaneous administration of physiological saline using an Alzet pump for 4 weeks.

[0269]

[0388] Furthermore, animals in groups 3 and 4 were subcutaneously administered SNF472 (Na6IP6, free base: 600 g / mol) sodium salt (free base 696.27 g / mol) at doses of 30 mg / kg and 45 mg / kg, respectively, twice daily. SNF472 was administered subcutaneously in physiological saline (2 mL / kg) twice daily from D1 to D21. Animals in group 5 were subcutaneously administered SNF472 dissolved in physiological saline at a dose of 400 mg / 4 weeks using an Alzet pump for 4 weeks.

[0270]

[0389] In the animals of group 6, cilostazol (C) in 5% CMC sodium salt (5 mL / kg) aqueous solution was administered. 20 H 27 N5O2, free base 369.46 g / mol; lot number LRAB9590, Sigma-Aldrich Corp., St. Louis, MO, US) 45 mg / kg was administered orally daily from D1 to D21.

[0271]

[0390] On the days when hemoperfusion tests were performed with the use of effective drugs (i.e., D10, D17, and D21), the following doses were administered:

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

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

[0393] Under the above scheme, effective drugs are identified by their maximum plasma concentration (C) at the time the test measurement is performed. max ) should be the case.

[0272]

[0394] Limb ischemia was assessed in all rats during treatment (i.e., D0, D10, D17, and D21) by laser Doppler perfusion imaging. 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) with (a) Groups 2a and 2b (placebo), (b) Groups 3, 4, and 5 (SNF472), and (c) Group 6 (cilostazol).

[0273]

[0395] SNF472 attenuated limb ischemia in uremic rats in a dose-response manner. Treatment with cilostazol alone was ineffective in this model. 3. Calcium content and calcification - ICP-OES

[0396] The subjects were anesthetized, and their blood was collected on day 21. The subjects were euthanized after blood loss. They were then autopsied, and their hearts and aortic arteries were collected. The tissues were freeze-dried for 24 hours and weighed. The freeze-dried tissues were then immersed in a dry bath incubator with a 1:1 HNO3:HClO4 mixture at 180°C for 2-4 hours. Subsequently, the immersed tissues were diluted to a final volume of 10 mL using ultra-high purity Milli-Q water (MilliporeSigma (Merck KGaA), Burlington, MA, US). The calcium content in the tissue samples was quantified by ICP-OES.

[0274]

[0397] SNF472, compared to placebo, was shown to be effective against cardiac and vascular calcification in uremic rats, for example, in the cardiac and aortic artery, after daily subcutaneous administration and / or delivery by Alzet pump, in a dose-response manner. Cilostazol was not active against calcification at the same dose. The following is a description of the claims as they were at the time of filing the application. [Claim 1] Compounds of general formula I, or pharmaceutically acceptable salts thereof, for use in increasing tissue perfusion and / or oxygen supply in subjects requiring such perfusion and / or oxygen supply. [C1] JPEG0007919861000011.jpg4665[wherein, R 1、 R 3、 R 5、 R 7、 R9 and R 11 These are independently OH, the base of formulas II, III, and IV: [C2] Selected from JPEG0007919861000012.jpg44141 and parts of a different structure, however, (i) R1, R3, R5, R7, R9 and R 11 At least one of these is selected from the bases of equations II, III, and IV, (ii)R 1、 R 3、 R 5、 R 7、 R9 and R11 [The condition is that 0, 1, 2, or 3 of these are parts of a different structure.] [Claim 2] Compounds of general formula I, or pharmaceutically acceptable salts thereof, for use in the treatment or prevention of ischemia and / or ischemia-related diseases or conditions in subjects requiring such treatment or prevention. [C3] JPEG0007919861000013.jpg5055[In the formula, R1, R3, R5, R7, R9 and R 11 These are independently OH, the base of formulas II, III, and IV: [C4] Selected from JPEG0007919861000014.jpg45140 and parts of a different structure, however, (i) R1, R3, R5, R7, R9 and R 11 At least one of these is selected from the bases of equations II, III, and IV, (ii)R 1、 R 3、 R 5、 R 7、 R9 and R 11 [The condition is that 0, 1, 2, or 3 of these are parts of a different structure.] [Claim 3] The heterogeneous structural parts are the base of formula V, the base of formula VI, and the base of formula VII: [5] JPEG0007919861000015.jpg80131[In the formula, n is an integer in the range of 2 to 200, R 13 A compound for use according to claim 1 or 2, selected from H, methyl, and ethyl. [Claim 4] A compound for use according to any one of claims 1 to 3, for the treatment or prevention of peripheral artery disease. [Claim 5] A compound for use according to any one of claims 1 to 4, for the treatment or prevention of severe limb ischemia. [Claim 6] A compound for use according to any one of claims 1 to 5, which is a sodium salt. [Claim 7] A compound for use according to any one of claims 1 to 6, wherein the compound of formula I is inositol hexaphosphate. [Claim 8] The compound for use according to claim 7, wherein inositol hexaphosphate is myo-inositol hexaphosphate. [Claim 9] A compound for use according to claim 7 or 8, which is a hexasodium salt. [Claim 10] R1, R3, R5, R7, R9 and R 11 A compound for use according to any one of claims 1 to 6, wherein one or two of the groups are selected from the groups of formulas V, VI, and VII. [Claim 11] R1, R5, R9 and R 11 The compound for use according to claim 10, wherein R3 and R7 are groups of formula II, and R3 and R7 are groups of formula V. [Claim 12] The basis of equation V is that n is in the range of 2 to 200, and R 13 The compound for use according to claim 11, wherein H is present. [Claim 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 pharmaceutically acceptable excipients and carriers. [Claim 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. [Claim 15] 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 undergoing dialysis. [Claim 16] A compound or pharmaceutical composition for use according to claim 15, administered to a subject undergoing hemodialysis. [Claim 17] A compound or pharmaceutical composition for use according to claim 16, which is administered to unfiltered blood extracted from a subject. [Claim 18] A compound for use according to any one of claims 1 to 12 or a pharmaceutical composition for use according to claim 13, administered via a parenteral route. [Claim 19] The compound or pharmaceutical composition for use according to claim 18, wherein parenteral administration is intravenous, subcutaneous, or intramuscular. [Claim 20] A compound of general formula I according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 13, for use in improving the walking ability of subjects requiring such improvement. [Claim 21] A compound of general formula I according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 13, for use in increasing the maximum walking distance (MWD), maximum walking time (MWT), or both, in subjects for which it is necessary to increase those. [Claim 22] A compound or pharmaceutical composition for use according to any one of claims 1 to 22, wherein the compound is administered to a subject in an effective dose of about 0.001 mg / kg to about 60 mg / kg.

Claims

1. A pharmaceutical composition for increasing tissue perfusion and / or oxygen supply, comprising myo-inositol hexaphosphate or a pharmaceutically acceptable salt thereof, for the treatment or prevention of peripheral artery disease, and The pharmaceutical composition does not contain a cationic, lipophilic, water-soluble molecule, and the cationic, lipophilic, water-soluble molecule contains at least one cationic functional group selected from the group consisting of guanidium, imidazolium, 1,2-diammonium ethylene, 1,8-diammonium naphthyl, and 2,2'-bipyridinium. The aforementioned pharmaceutical composition.

2. A pharmaceutical composition comprising myo-inositol hexaphosphate or a pharmaceutically acceptable salt thereof for the treatment or prevention of ischemia and / or ischemia-related disease or condition, for the treatment or prevention of peripheral artery disease, and The pharmaceutical composition does not contain a cationic, lipophilic, water-soluble molecule, and the cationic, lipophilic, water-soluble molecule contains at least one cationic functional group selected from the group consisting of guanidium, imidazolium, 1,2-diammonium ethylene, 1,8-diammonium naphthyl, and 2,2'-bipyridinium. The aforementioned pharmaceutical composition.

3. A pharmaceutical composition according to claim 1 or 2, for the treatment or prevention of severe limb ischemia.

4. The pharmaceutical composition according to any one of claims 1 to 3, comprising the sodium salt of myo-inositol hexaphosphate.

5. A pharmaceutical composition according to any one of claims 1 to 4, comprising the hexasodium salt of myo-inositol hexaphosphate.

6. A pharmaceutical composition as defined in any one of claims 1 to 3, further comprising pharmaceutically acceptable excipients and carriers.

7. A pharmaceutical composition according to any one of claims 1 to 6, to be administered to a subject with renal failure.

8. A pharmaceutical composition according to any one of claims 1 to 6, to be administered to a subject undergoing dialysis.

9. The pharmaceutical composition according to claim 8, administered to a subject undergoing hemodialysis.

10. The pharmaceutical composition according to claim 9, which is administered to unfiltered blood extracted from the subject.

11. A pharmaceutical composition according to any one of claims 1 to 6, administered via a parenteral route.

12. The pharmaceutical composition according to claim 11, wherein parenteral administration is intravenous, subcutaneous, or intramuscular administration.

13. A pharmaceutical composition according to any one of claims 1 to 6, for improving the walking ability of a patient who has peripheral artery disease or is at risk of developing such disease.

14. A pharmaceutical composition according to any one of claims 1 to 6 for increasing the maximum walking distance (MWD), maximum walking time (MWT), or both, in a patient having peripheral artery disease or at risk of developing such disease.

15. A pharmaceutical composition according to any one of claims 1 to 14, wherein the myo-inositol hexaphosphate or a pharmaceutically acceptable salt thereof is administered in an effective dose of about 0.001 mg / kg to about 60 mg / kg.

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