Treatment methods for cholesterol crystal embolism using cyclodextrins

Abstract

Disclosed herein are methods for reducing the amount and / or size of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) in an individual, and / or methods for altering the shape of such crystals (and / or blood clots). Further disclosed herein are methods for treating cholesterol crystal embolism (CCE) and / or symptoms thereof in an individual. The methods generally comprise administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual. Further provided herein are pharmaceutical compositions comprising a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin and a pharmaceutically acceptable excipient.

Description

Technical Field

[0001] Cross-reference This application claims the benefit of U.S. Provisional Application No. 63 / 071,222, filed Aug. 27, 2020, which is hereby incorporated by reference in its entirety.

Background Art

[0002] Cholesterol crystal embolism (CCE) (also called "atheroembolism", "atheroembolic syndrome", or "cholesterol embolism syndrome") is a systemic disease thought to result from the dissemination of cholesterol crystals from large blood vessels (e.g., the aorta), often due to the rupture of atherosclerotic plaques, into the peripheral circulation, and the occlusion of smaller arteries and other blood vessels by these crystals. CCE has a variety of clinical manifestations, including symptoms in the kidneys, skin, central nervous system, intestine, and eyes. CCE is most commonly considered iatrogenic, such as a complication of a medical procedure involving blood vessels. However, in some patients (e.g., those with advanced atherosclerosis), CCE may occur spontaneously. There are no known treatments that have been shown to alter the prognosis of CCE. Preclinical data suggest that 2-hydroxypropyl-β-cyclodextrin may have important and beneficial effects on the pathological mechanisms underlying CCE by reducing the amount and / or size of circulating cholesterol crystals (and / or thrombi containing cholesterol crystals) and / or altering the shape of said crystals (and / or thrombi). Thus, 2-hydroxypropyl-β-cyclodextrin may offer a novel treatment option for CCE and its symptoms.

Summary of the Invention

[0003] Effective treatment of cholesterol crystal embolism (CCE) and / or its symptoms is needed. The present disclosure addresses this unmet need.

[0004] In one embodiment, a method is provided for reducing the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in an individual, for reducing the size of the crystals and / or blood clots, and / or for altering the shape of the crystals and / or blood clots, the method comprising administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual to reduce the amount of circulating cholesterol crystals in the individual, reduce the size of the crystals, and / or alter the shape of the crystals. In some cases, the size of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or more than 50% compared to the size of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals before administration of 2-hydroxypropyl-β-cyclodextrin. In some cases, the size of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals is the average size or the maximum size. In some cases, the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or more than 50% compared to the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals before administration of 2-hydroxypropyl-β-cyclodextrin. In some cases, the amount of circulating cholesterol crystals is the concentration of circulating cholesterol crystals. In some cases, the circulating cholesterol crystals are present in the blood, plasma, or serum. In some cases, treatment reduces the incidence or severity of cholesterol crystal embolism (CCE) in the individual.

[0005] In another embodiment, a method for treating cholesterol crystal embolism (CCE) and / or one or more of its symptoms in an individual is provided, comprising administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual, thereby treating the cholesterol crystal embolism (CCE) and / or one or more of its symptoms in the individual. Optionally, the treatment includes reducing the size of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in the individual. Optionally, the size of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals may be the maximum size or the average size. Optionally, the treatment includes reducing the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in the individual. Optionally, the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals may be the concentration of circulating cholesterol crystals. Optionally, the treatment includes altering the shape of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals in the individual. Optionally, the treatment includes reducing inflammation in the individual. Optionally, the inflammation may be determined by cytokine protein levels and / or RNA levels. Depending on the circumstances, the above treatment may include improving renal function in the individual. Depending on the circumstances, the above treatment may include improving dermatological symptoms in the individual. Depending on the circumstances, the above treatment may include improving eosinophilia in the individual. Depending on the circumstances, the above treatment may include improving hematological abnormalities in the individual. Depending on the circumstances, the above treatment may include improving complement levels in the individual. Depending on the circumstances, the above treatment may include improving proteinuria in the individual.

[0006] Depending on the circumstances, the effective therapeutic dose is approximately 50 mg / kg to approximately 2,500 mg / kg. Depending on the circumstances, the effective therapeutic dose is approximately 4 g to approximately 250 g. Depending on the circumstances, the effective therapeutic dose is sufficient to achieve serum, plasma, and / or whole blood concentrations of approximately 0.01 mM to approximately 3 mM of 2-hydroxypropyl-β-cyclodextrin. Depending on the circumstances, the effective therapeutic dose is sufficient to increase the circulating and / or systemic levels of one or more oxysterols in the individual by at least approximately 10% compared to before administration. Depending on the circumstances, the one or more oxysterols may be 24S-hydroxycholesterol, 27-hydroxycholesterol, or both. Depending on the circumstances, the effective therapeutic dose is sufficient to increase the cholesterol crystal solubility (CCDC) of plasma by at least approximately 10% compared to before administration. In some cases, the therapeutically effective dose is an amount effective in increasing ABCA1 and / or ABCG1 mRNA levels by at least about 10% after administration compared to before administration. In some cases, the 2-hydroxypropyl-β-cyclodextrin is selected from the group consisting of Kleptose® HP parenteral grade, Kleptose® HPB parenteral grade, Kleptose® HPB-LB parenteral grade, Cavitron® W7 HP5 Pharma cyclodextrin, Cavitron® W7 HP7 Pharma cyclodextrin, Trappsol® Cyclo®, and VTS-270 / Adorabetadex. In some cases, the individual has one or more risk factors for CCE. In some cases, one or more of the above risk factors for CCE are selected from the group consisting of interventional vascular procedures, interventional diagnostic procedures, cardiovascular surgery, cardiovascular disease (e.g., coronary artery disease, atherosclerotic cardiovascular disease), aortic aneurysm, aortic plaque, hypertension, diabetes mellitus, hyperlipidemia, smoking, being male, age, increased inflammation (e.g., elevated serum (hs)CRP levels), anticoagulant therapy, thrombolytic therapy, and any combination thereof. In some cases, the above individuals have obtained one or more analytical test results associated with CCE.Depending on the circumstances, one or more analytical test results associated with the above CCE may be selected from the group consisting of elevated serum creatinine, leukocytosis, eosinophilia, anemia, thrombocytopenia, hypocomplementemia, elevated erythrocyte sedimentation rate, elevated (hs)CRP levels, elevated fibrinogen levels, eosinophilia, proteinuria, hematuria, abnormal liver enzymes, and any combination thereof. Depending on the circumstances, the individual may be at least 30 years old. Depending on the circumstances, the individual may be human. Depending on the circumstances, the administration may further include (i) administering to the individual a therapeutically effective first dose of 2-hydroxypropyl-β-cyclodextrin at a first time point, and (ii) administering to the individual a therapeutically effective second dose of 2-hydroxypropyl-β-cyclodextrin at a second time point. Depending on the circumstances, the second time point may be at least 4 hours, at least 6 hours, at least 8 hours, at least 12 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the first time point. Depending on the circumstances, the above administration may be done by intravenous administration.

[0007] In another embodiment, a pharmaceutical composition is provided comprising 2-hydroxypropyl-β-cyclodextrin in an amount effective for reducing the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in an individual, reducing the size of the crystals and / or blood clots, and / or altering the shape of the crystals and / or blood clots, and a pharmaceutically acceptable excipient. In yet another embodiment, a pharmaceutical composition is provided comprising 2-hydroxypropyl-β-cyclodextrin in an amount effective for treating cholesterol crystal embolism (CCE) and / or its symptoms in an individual, and a pharmaceutically acceptable excipient. Optionally, the amount of 2-hydroxypropyl-β-cyclodextrin is effective for increasing the circulating level and / or systemic level of one or more oxysterols in the individual by at least about 10% after administration of the pharmaceutical composition to the individual. Optionally, the one or more oxysterols are 24S-hydroxycholesterol, 27-hydroxycholesterol, or both. In some cases, the amount of 2-hydroxypropyl-β-cyclodextrin is effective in increasing the plasma cholesterol crystal solubility (CCDC) of the individual by at least about 10% after administration of the pharmaceutical composition to the individual. In some cases, the amount of 2-hydroxypropyl-β-cyclodextrin is effective in increasing the mRNA levels of ABCA1 and / or ABCG1 of the individual by at least about 10% after administration of the pharmaceutical composition to the individual. In some cases, the pharmaceutical composition is formulated for single-dose administration. In some cases, the pharmaceutical composition is formulated for intravenous administration.

[0008] In yet another embodiment, a kit is provided comprising (a) one or more containers and (b) any one of the aforementioned pharmaceutical compositions contained within the one or more containers. Optionally, the kit further comprises (c) instructions for the use of the pharmaceutical composition to reduce the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in an individual, to reduce the size of the crystals and / or blood clots, and / or to alter the shape of the crystals and / or blood clots, and / or to treat CCE or one or more of its symptoms in an individual. Optionally, at least one of the one or more containers is an intravenous infusion bag. Optionally, the one or more containers comprise a single container containing the pharmaceutical composition and one or more further active pharmaceutical ingredients. Optionally, the one or more containers comprise a first container containing the pharmaceutical composition and a second container containing one or more further active pharmaceutical ingredients. Optionally, the kit further comprises one or more further components selected from the group consisting of intravenous infusion bags, catheters, tubes, needles, syringes, solutions, and any combination thereof.

[0009] In another embodiment, a method is provided for reducing the amount and / or size of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) in an individual, and / or for altering the shape of the crystals (and / or blood clots), the method comprising administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual to reduce the amount and / or size of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) in the individual, and / or altering the shape of the crystals (and / or blood clots). In some cases, the size (e.g., average size, maximum size) of the circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) is reduced by at least about 10% (e.g., at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% or more) compared to the size (e.g., average size, maximum size) of the circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the amount (e.g., concentration) of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) is reduced by at least about 10% (e.g., at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%) compared to the amount (e.g., concentration) of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the above treatment reduces the incidence or severity of cholesterol crystal embolism (CCE) in the above individual.

[0010] In another embodiment, a method for treating cholesterol crystal embolism (CCE) and / or one or more of its symptoms in an individual is provided, comprising administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual, thereby treating the CCE and / or one or more of its symptoms in the individual. Optionally, the treatment may include reducing the size and / or volume of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) in the subject. In some cases, the size (e.g., average size, maximum size) of the circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) is reduced by at least about 10% (e.g., at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% or more) compared to the size (e.g., average size, maximum size) of the circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the amount (e.g., concentration) of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) is reduced by at least about 10% (e.g., at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%) compared to the amount (e.g., concentration) of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the treatment alters the shape of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals). In some cases, the treatment reduces inflammation (as determined by, for example, cytokine protein levels and / or RNA levels) compared to the level of inflammation before treatment with 2-hydroxypropyl-β-cyclodextrin.In some cases, the above treatment improves renal function compared to renal function before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the above treatment improves dermatological symptoms compared to before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the above treatment improves eosinophilia compared to before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the above treatment improves hematological abnormalities compared to before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the above treatment improves complement levels compared to before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the above treatment improves proteinuria compared to before treatment with 2-hydroxypropyl-β-cyclodextrin.

[0011] In any one of the prior art embodiments, the effective therapeutic dose is approximately 50 mg / kg to approximately 2,500 mg / kg. In any one of the prior art embodiments, the effective therapeutic dose is approximately 4 g to approximately 250 g. In any one of the prior art embodiments, the effective therapeutic dose is sufficient to achieve serum, plasma, and / or whole blood concentrations of approximately 0.01 mM to approximately 3 mM of 2-hydroxypropyl-β-cyclodextrin. In any one of the prior art embodiments, the 2-hydroxypropyl-β-cyclodextrin is selected from the group consisting of Kleptose® HP parenteral grade, Kleptose® HPB parenteral grade, Kleptose® HPB-LB parenteral grade, Cavitron® W7 HP5 Pharma cyclodextrin, Cavitron® W7 HP7 Pharma cyclodextrin, Trappsol® Cyclo®, and VTS-270 / Adrabetadex. In any one of the preceding embodiments, the subject has one or more risk factors for CCE. These one or more risk factors for CCE may be selected from the group consisting of interventional vascular procedures, interventional diagnostic procedures, cardiovascular surgery, cardiovascular disease (e.g., coronary artery disease, atherosclerotic cardiovascular disease), aortic aneurysm, aortic plaque, hypertension, diabetes mellitus, hyperlipidemia, smoking, being male, age, increased inflammation (e.g., elevated serum (hs)CRP levels), anticoagulation therapy, and thrombolytic therapy. In any one of the preceding embodiments, the subject has obtained one or more analytical test results associated with CCE. These one or more analytical test results associated with CCE may be selected from the group consisting of elevated serum creatinine, leukocytosis, eosinophilia, anemia, thrombocytopenia, hypocomplementemia, elevated erythrocyte sedimentation rate, elevated (hs)CRP levels, elevated fibrinogen levels, eosinophilia, proteinuria, hematuria, and abnormal liver enzymes. In any one of the preceding embodiments, the individual is at least 30 years old (for example, at least 40, at least 50, at least 60, at least 70, at least 80, or at least 90 years old). In any one of the preceding embodiments, the individual is human.In any one of the preceding embodiments, the administration further comprises (i) administering to the individual a therapeutically effective first dose of 2-hydroxypropyl-β-cyclodextrin at a first time point, and (ii) administering to the individual a therapeutically effective second dose of 2-hydroxypropyl-β-cyclodextrin at a second time point. In any one of the preceding embodiments, the second time point is at least 4 hours, at least 6 hours, at least 8 hours, at least 12 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the first time point. In any one of the preceding embodiments, the administration is by intravenous administration.

[0012] In another embodiment, a pharmaceutical composition is provided comprising 2-hydroxypropyl-β-cyclodextrin in an amount effective for reducing the amount and / or size of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) in an individual, and / or altering the shape of the crystals (and / or blood clots), and a pharmaceutically acceptable excipient.

[0013] In another embodiment, a pharmaceutical composition is provided comprising 2-hydroxypropyl-β-cyclodextrin in an amount effective for treating cholesterol crystal embolism (CCE) and / or its symptoms in an individual, and a pharmaceutically acceptable excipient.

[0014] In any one of the prior embodiments, the pharmaceutical composition is formulated for single-dose administration. In any one of the prior embodiments, the pharmaceutical composition is formulated for intravenous administration.

[0015] support All publications, patents, and patent applications referenced herein are incorporated herein by reference in the same way that each individual publication, patent, or patent application is specifically and individually indicated to be incorporated herein by reference.

[0016] Novel features of this disclosure are described in detail in the attached claims. The features and advantages of this disclosure will be better understood by referring to the following detailed description, which describes exemplary embodiments in which the principles of this disclosure are utilized, and to the attached drawings below. [Brief explanation of the drawing]

[0017] [Figure 1A] This figure shows a non-limiting example of the cholesterol crystal lysis capacity (CCDC) of plasma obtained from male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to an embodiment of the present disclosure. [Figure 1B] This figure shows a non-limiting example of cholesterol crystal lysis capacity (CCDC) of plasma obtained from male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to embodiments of the present disclosure. [Figure 2A] This figure shows a non-limiting example of total cholesterol levels in plasma obtained from male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to embodiments of the present disclosure. [Figure 2B] This figure shows a non-limiting example of plasma oxysterol levels obtained from male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to embodiments of the present disclosure. [Figure 2C] This figure shows a non-limiting example of plasma oxysterol levels obtained from male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to embodiments of the present disclosure. [Figure 3A] This figure shows a non-limiting example of mRNA levels of the gene ABCA1, which is regulated by the LXR transcription factor, in male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to embodiments of the present disclosure. [Figure 3B]This figure shows a non-limiting example of mRNA levels of the LXR transcription factor-regulated gene ABCG1 in male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to embodiments of the present disclosure. [Figure 3C] This figure shows a non-limiting example of mRNA levels of the gene ABCA1, which is regulated by the LXR transcription factor, in male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to embodiments of the present disclosure. [Figure 3D] This figure shows a non-limiting example of mRNA levels of the LXR transcription factor-regulated gene ABCG1 in male human subjects treated with gradually increasing doses of 2-hydroxypropyl-β-cyclodextrin according to embodiments of the present disclosure. [Modes for carrying out the invention]

[0018] This specification discloses methods for reducing the amount and / or size of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) in an individual (e.g., a human), and / or for altering the shape of the crystals (and / or blood clots). Optionally, the methods include treating cholesterol crystal embolism (CCE) (e.g., by preventing occlusion of small vessels by cholesterol crystal embolisms, cholesterol crystal clots, cholesterol crystal / protein clots, cholesterol crystal / DNA clots (e.g., extracellular traps)). Optionally, the methods include treating one or more symptoms and / or clinical manifestations of CCE. Optionally, the methods include treating ischemia of various organs and / or tissues resulting from CCE. Generally speaking, the methods provided herein involve administering a therapeutically effective amount of cyclodextrin to a subject in need (e.g., a subject with high levels of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals)). In certain embodiments, the cyclodextrin is 2-hydroxypropyl-β-cyclodextrin.

[0019] In some embodiments, methods for preventing the formation of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) in an individual (e.g., human) or for reducing the risk of such formation. Further disclosures include methods for preventing an increase in the amount and / or size of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) in an individual (e.g., human) or for reducing the risk of such an increase, and / or for altering the shape of such crystals (and / or blood clots). Optionally, the methods include preventing or reducing the risk of developing cholesterol crystal embolism (CCE) (for example, by preventing or reducing the risk of occlusion of small vessels by cholesterol crystal embolisms, cholesterol crystal clots, cholesterol crystal / protein clots, cholesterol crystal / DNA clots (e.g., extracellular traps)). Optionally, the methods include preventing or reducing the risk of developing symptoms and / or clinical symptoms of CCE. Depending on the circumstances, the above methods may include preventing or reducing the risk of ischemia of various organs and / or tissues resulting from, for example, CCE (and / or its symptoms or clinical manifestations, such as renal injury, atherothrombotic renal disease (ARD)). In general terms, the methods provided herein include administering a therapeutically effective amount of cyclodextrin to a subject in need (for example, prophylactically, to a subject at risk of developing CCE). In certain embodiments, the cyclodextrin is 2-hydroxypropyl-β-cyclodextrin.

[0020] In some embodiments, methods of treating cardiovascular disease are disclosed herein. In some cases, the cardiovascular disease is an atherosclerotic cardiovascular disease (e.g., a cardiovascular disease caused by or involving atherosclerosis). The atherosclerotic cardiovascular disease may be any one of coronary artery disease (CAD), stroke, peripheral artery disease (PAD), peripheral vascular disease (PVD), chronic kidney disease (CKD) caused by atherosclerosis, end-stage kidney disease (ESKD) caused by atherosclerosis, acute kidney injury caused by atherosclerosis, atherosclerotic renovascular disease (ARVD), renal artery stenosis, aortic aneurysm, idiopathic peripheral atrial hypertension, erectile dysfunction, intermittent claudication, and / or postoperative or iatrogenic arterial disease. In some cases, PAD includes lower extremity arterial disease. In some cases, the method includes treating and / or preventing atherosclerosis. In some cases, the method includes treating a subject suspected of having or at risk of developing acute coronary syndrome (ACS) or chronic coronary syndrome (CCS) (e.g., as defined by the European Society of Cardiology), or having ACS or CCS. In some aspects, the method may include administering a therapeutically effective amount of cyclodextrin to a subject that needs it (e.g., a subject having a cardiovascular disease (e.g., an atherosclerotic cardiovascular disease), suspected of having a cardiovascular disease, or at risk of developing a cardiovascular disease). In some cases, the therapeutically effective amount is an amount effective to increase the circulating and / or systemic levels of one or more sterols and / or oxysterols in the subject as compared to a baseline (e.g., prior to treatment with cyclodextrin). In certain aspects, the cyclodextrin is 2-hydroxypropyl-β-cyclodextrin.

[0021] For the following terms, in addition to the understanding of these terms by those skilled in the art, the meaning of these terms used in this specification will be discussed to explain. In this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. It should be further noted that the claims can be drafted to exclude any optional elements. Thus, this description is intended to serve as a prerequisite for the use of exclusive terms such as "alone", "only", etc. in relation to the description of the components of the claims or the use of "negative" limitations.

[0022] In this specification, the term "about" + a number refers to that number ± 10% of that number. The term "about" + a range refers to that range - 10% of the minimum value and + 10% of the maximum value.

[0023] In this specification, the terms "subject", "individual", and "patient" are used synonymously. None of these terms should be construed as requiring the management of medical professionals (e.g., physicians, nurses, physician assistants, nursing assistants, hospice staff). In this specification, a subject can be any animal, including mammals (e.g., humans or non-human animals) and non-mammals. In one embodiment, the subject is a human.

[0024] In this specification, the terms “to treat,” “to treat,” or “treatment,” and other grammatical equivalents include: improving or preventing the underlying cause of one or more symptoms of a disease or illness; alleviating, reducing, or improving one or more symptoms of a disease or illness; improving, preventing, or reducing the appearance, severity, or frequency of one or more symptoms of a disease or illness; suppressing a disease or illness, for example, preventing the onset of a disease or illness, alleviating a disease or illness, causing regression of a disease or illness, alleviating a condition caused by a disease or illness, or suppressing the symptoms of a disease or illness in a preventive and / or therapeutic manner. The treatment methods disclosed herein include disclosures of the use of (e.g., pharmaceutical) compositions provided herein for the treatment of any indication described herein.

[0025] The term "pharmaceutically acceptable" generally refers to the attributes of a material useful for preparing a pharmaceutical composition that is safe, non-toxic, and not biologically or otherwise undesirable, and is acceptable for veterinary and human pharmaceutical use. "pharmaceutically acceptable" may also refer to a material such as a carrier or diluent that does not inhibit the biological activity or properties of the compound and is relatively non-toxic, for example, that can be administered to an individual without causing undesirable biological effects or interacting in a harmful manner with any of the components of the composition containing the material.

[0026] As used herein, "pharmaceutically acceptable excipients" means any pharmaceutically acceptable component in a pharmaceutical composition that is non-therapeutic and non-toxic to the recipient, such as disintegrants, binders, bulking agents, solvents, buffers, isotonic agents, stabilizers, antioxidants, surfactants, carriers, diluents, excipients, preservatives, or lubricants used in the formulation of pharmaceuticals.

[0027] As used herein, the terms “effective dose” or “therapeutic effective dose” mean a sufficient amount of an agent or compound administered to alleviate, to some extent, one or more symptoms of the disease or illness being treated, or to reduce the underlying cause of the disease or illness being treated. In some embodiments, the result is a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or any other desired change in the biological system. For example, “effective dose” for therapeutic use is the amount of a composition containing the compound disclosed herein required to clinically significant reduce the symptoms or underlying causes of the disease (e.g., without excessive adverse side effects). In some embodiments, the appropriate “effective dose” in any individual case is determined using techniques such as dose-escalation studies. The term “therapeutic effective dose” includes, for example, a prophylactic effective dose. “Effective dose” of a compound disclosed herein may be an amount effective to achieve the desired effect or therapeutic improvement (e.g., without excessive adverse side effects). “Effective dose” of a compound disclosed herein may be an amount effective to achieve one or more desired outcomes. It is important to understand that, in some cases, the “effective dose” or “therapeutic effective dose” may vary from subject to subject due to differences in the metabolism of the composition, the subject’s age, weight, general condition, any concomitant medications the subject may be taking, the disease being treated, the severity of the disease being treated, and the prescribing physician’s judgment. In some cases, the disease or condition being treated is CCE. In some cases, the disease or condition being treated is a disease or condition related to or caused by CCE.

[0028] Treatment methods for cholesterol crystal embolism and its symptoms

[0029] Examples 1-3 of this specification demonstrate increased plasma cholesterol crystal lysis capacity, elevated oxysterol levels, and elevated mRNA levels of the LXR transcription factor-regulated genes ABCA1 and ABCG1 in human subjects treated with 2-hydroxypropyl-β-cyclodextrin. These data suggest that 2-hydroxypropyl-β-cyclodextrin can be used to treat cholesterol crystal embolism (CCE) as described herein.

[0030] Accordingly, this specification discloses methods for treating subjects who have, are suspected of having, or are at risk of developing CCE or its symptoms and / or clinical manifestations (for example, by reducing the amount and / or size of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals), and / or altering the shape of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals). In some cases, the symptoms and / or clinical manifestations of CCE are one or more skin manifestations of CCE. These one or more skin manifestations of CCE include, but are not limited to, reticular dermatosis (e.g., purple discoloration of the skin), cyanosis (e.g., bluish discoloration of the skin due to poor blood circulation or oxygen deficiency), gangrene (e.g., death of body tissue due to lack of blood flow), skin ulcers, purpura, erythematous nodules, and blue toe syndrome. In some cases, the symptoms or clinical manifestations of CCE are atherothrombotic nephropathy. In some cases, the symptoms or clinical manifestations of CCE described above are one or more renal symptoms of CCE. These one or more renal symptoms of CCE include, but are not limited to, acute kidney injury, subacute kidney injury, chronic kidney injury, malignant hypertension, glomerulonephritis, end-stage renal disease, allograft dysfunction, and renal infarction. In some cases, the symptoms or clinical manifestations of CCE described above are one or more gastrointestinal symptoms of CCE. These one or more gastrointestinal symptoms of CCE include, but are not limited to, abdominal pain, diarrhea, bleeding, intestinal ischemia, intestinal infarction, intestinal perforation, necrotizing pancreatitis, focal hepatocyte necrosis, and acalculous cholecystitis. In some cases, the symptoms or clinical manifestations of CCE described above are one or more central nervous system symptoms of CCE. These one or more central nervous system symptoms of CCE include, but are not limited to, headache, dizziness, confusion, amnesia, transient ischemic attack, stroke, cerebral infarction, spinal cord infarction, paraplegia, and mononeuropathy. In some cases, the symptoms or clinical manifestations of CCE described above are one or more ocular symptoms of CCE. These one or more ocular symptoms of CCE include, but are not limited to, transient monocular blindness, eye pain, blurred vision, and Hollenhorst spots. In some cases, the symptoms or clinical manifestations of CCE described above are one or more of the following: myocardial infarction, adrenal insufficiency, penile necrosis, myositis, rhabdomyolysis, splenic infarction, and alveolar hemorrhage.In some cases, the symptoms or clinical manifestations of CCE may include, but are not limited to, one or more of the following: fever, fatigue, loss of appetite, weight loss, and muscle pain.

[0031] In some cases, treating the subject as described herein reduces the size (e.g., average size, maximum size) of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) in the subject. In some cases, the size (e.g., average size, maximum size) of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) is reduced by at least about 10% (e.g., at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50% or more) compared to the size (e.g., average size, maximum size) of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, treating the subject as described herein reduces the amount (e.g., concentration) of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) in the subject. In some cases, the amount (e.g., concentration) of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) decreases by at least about 10% (e.g., at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%) compared to the amount (e.g., concentration) of circulating (e.g., blood, serum, plasma) cholesterol crystals (and / or blood clots containing cholesterol crystals) before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, treating the subject as described herein leads to the dissolution of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) in the subject. In some cases, treating the subject as described herein alters the shape of the circulating cholesterol crystals (and / or blood clots containing cholesterol crystals).In some cases, reducing the number and / or size of circulating cholesterol crystals (and / or cholesterol-containing blood clots) in the subject, and / or altering the shape of the crystals (and / or blood clots), improves or reduces CCE in the subject. In some cases, reducing the number and / or size of circulating cholesterol crystals (and / or cholesterol-containing blood clots) in the subject, and / or altering the shape of the crystals (and / or blood clots), improves or reduces one or more symptoms and / or clinical symptoms of CCE in the subject. In some cases, treating the subject as described herein reduces inflammation (determined, for example, by cytokine protein levels and / or RNA levels) compared to the level of inflammation prior to treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, treating the subject as described herein improves renal function compared to the level of renal function prior to treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, treating the subject as described herein improves dermatological symptoms compared to prior to treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, treating the subject as described herein improves eosinophilia compared to before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, treating the subject as described herein improves hematological abnormalities compared to before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, treating the subject as described herein improves complement levels compared to before treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, treating the subject as described herein improves proteinuria compared to before treatment with 2-hydroxypropyl-β-cyclodextrin.

[0032] In various embodiments, the above method involves administering cyclodextrin to a subject (who has, is suspected of having, or is at risk of developing, one or more diseases or illnesses related to CCE and / or CCE). Cyclodextrins are a family of cyclic oligosaccharides consisting of a ring (e.g., a macroring) of glucose subunits linked by α-1,4 glycosidic bonds. Cyclodextrins contain a large number of glucose monomers in their ring structure. Common cyclodextrins include α-cyclodextrin (consisting of 6 glucose monomers), β-cyclodextrin (consisting of 7 glucose monomers), γ-cyclodextrin (consisting of 8 glucose monomers), and δ-cyclodextrin (consisting of 9 glucose monomers). The outer portion of the ring structure is hydrophilic, and the internal cavity of the ring structure is hydrophobic; therefore, cyclodextrins are generally water-soluble (e.g., due to the hydrophilic outer portion) and can incorporate hydrophobic molecules into the cavity (e.g., due to the hydrophobic cavity). The water solubility of parent cyclodextrins is limited, and therefore several chemically modified cyclodextrins have been synthesized in which the hydroxyl group is replaced with another chemical moiety, for example, to improve solubility. In various embodiments, the methods provided herein involve administering a cyclodextrin to a subject (e.g., a human) that requires it (e.g., having an increased amount of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals); e.g., having CCE, suspected of having CCE, or at risk of developing CCE). In some cases, the subject has, is suspected of having, or is at risk of developing circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) (e.g., after the breakdown of atherosclerotic plaques), or has a high level of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals), or is suspected of having a high level, or is at risk of developing a high level.

[0033] In certain embodiments, the cyclodextrin is 2-hydroxypropyl-β-cyclodextrin. In some cases, the 2-hydroxypropyl-β-cyclodextrin is Kleptose® HP parenteral grade (Roquette Freres, #346114; accessible at roquette.com / - / media / roquette-sharepoint-libraries / sdol_product-specification-sheet / roquette_quality_specification-sheet_kleptose-hp-parenteral-grade_50_346114_en.pdf (as of August 26, 2020)), Kleptose® HPB parenteral grade (Roquette Freres, #346111; Accessible at roquette.com / - / media / roquette-sharepoint-libraries / sdol_product-specification-sheet / roquette_quality_specification-sheet_kleptose-hpb-parenteral-grade_50_346111_en.pdf (as of August 26, 2020)), Kleptose® HPB-LB parenteral grade (Roquette Freres, #346115; Accessible at roquette.com / - / media / roquette-sharepoint-libraries / sdol_product-specification-sheet / roquette_quality_specification-sheet_kleptose-hpb-lb-parenteral-grade_50_346115_en.pdf (as of August 26, 2020)), Cavitron® W7 HP5 Pharma Cyclodextrin (Ashland;ashland.com / file_source / Ashland / Product / Documents / Pharmaceutical / PC_l1734_Cavitron_Cavasol.The following are selected from the group consisting of (accessible as a PDF as of August 26, 2020): Cavitron® W7 HP7 Pharma cyclodextrin (accessible at Ashland; ashland.com / file_source / Ashland / Product / Documents / Pharmaceutical / PC_11734_Cavitron_Cavasol.pdf as of August 26, 2020), Trappsol® Cyclo® (Cyclo Therapeutics, Inc.; accessible at cyclotherapeutics.com / cyclodextrins / trappsol-cyclo as of August 26, 2020), and VTS-270 / Adorabetadex.

[0034] In certain embodiments, the cyclodextrins provided or used in (e.g., pharmaceutical) compositions, methods, or other applications herein are mixtures of multiple cyclodextrins, for example, in some embodiments, the 2-hydroxypropyl-β-cyclodextrin provided herein comprises a mixture of multiple 2-hydroxypropyl-β-cyclodextrins. In some embodiments, the cyclodextrin molecules provided herein are optionally substituted with one or more chemical groups, each independently being a hydroxypropyl group, a hydroxyethyl group, a methyl group, an ethyl group, a carboxymethyl group, a heptakis(2,6-di-O-methyl) group, a sulfoethyl group, a sulfopropyl group, and / or a sulfobutylethyl group, or are oligomers thereof. In some preferred embodiments, the cyclodextrin is a hydroxypropyl-β-cyclodextrin, such that one or more hydroxyls of the cyclodextrin are substituted with hydroxypropyl (e.g., a 2-hydroxypropyl group). For example, one or more hydroxyl positions are substituted with one or more hydroxypropyl groups by substituting the H of hydroxyl (OH) with a -CH2CH2(OH)CH3 group, as shown in Formula I below. In some embodiments, the 2-hydroxypropyl-β-cyclodextrin comprises a plurality of cyclodextrins having various different degrees of substitution (DS) and / or molar substitution (MS) values. [ka]

[0035] In some embodiments, the multiple β-cyclodextrin molecules in a β-cyclodextrin (a mixture of multiple β-cyclodextrin molecules) are characterized by average molar substitution. “Molar substitution,” or “MS,” is the average number of substituents per glucose unit in the multiple β-cyclodextrin molecules. In some embodiments, the MS is measured according to the procedure described in the USP monograph for hydroxypropyl betadex (USP NF 2015) ("USP Hydroxypropyl Betadex Monograph") (the entire monograph is incorporated herein by reference). In some embodiments, the (e.g., pharmaceutical) compositions provided herein contain multiple β-cyclodextrin molecules having an average MS of at least about 0.3. In some embodiments, the (e.g., pharmaceutical) compositions provided herein contain multiple β-cyclodextrin molecules having an average MS of about 0.3 to 1.0.

[0036] In some embodiments, the above-mentioned multiple types of β-cyclodextrin molecules are characterized by their mean degree of substitution. The term “degree of substitution,” or “DS,” refers to the total number of substituents directly or indirectly substituted on the β-cyclodextrin molecule. In some embodiments, the β-cyclodextrin molecule may have one or more glucose units substituted by substituents at the hydroxyl position. Thus, mean DS is the total number of substituents in the population of β-cyclodextrins divided by the number of β-cyclodextrin molecules. In some embodiments, the mean DS of the molecules is measured using electrospray ionization mass spectrometry (ESI-MS) analysis (e.g., HPLC-ESI-MS). In some embodiments, the mean DS of the molecules is measured by the peak height of the electrospray ionization MS spectrum. In some embodiments, the mean DS of the molecules is determined by multiplying the MS by 7. In some embodiments, the (e.g., pharmaceutical) compositions provided herein contain multiple types of β-cyclodextrin molecules having mean DS of about 2.0 to 7.0.

[0037] In some embodiments, any atom of the cyclodextrin described herein (e.g., 2-hydroxypropyl-β-cyclodextrin) may be substituted with any appropriate isotope. In certain embodiments, any one or more hydrogen atoms of the cyclodextrin described herein (e.g., 2-hydroxypropyl-β-cyclodextrin) may be substituted or replaced with deuterium atoms. Such cyclodextrins are expected to have properties similar to or improved compared to the original cyclodextrin without deuterium. Deuterium is a safe and stable non-radioactive isotope of hydrogen. Deuterium forms stronger bonds with carbon than hydrogen. In some cases, the increased bond strength conferred by deuterium positively affects the properties of the cyclodextrin, creating the potential to improve the efficacy, safety, and / or tolerability of the drug. Furthermore, deuteration may reduce in vivo metabolic clearance, thereby increasing the half-life and circulation of the compound. At the same time, since the size and shape of deuterium are essentially identical to those of hydrogen, it is expected that the substitution of hydrogen with deuterium will not affect the biochemical potency and selectivity of the compound compared to the original chemical substance containing only hydrogen.

[0038] In various embodiments, the subject is administered a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin. In some embodiments, administration of a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin increases the circulating and / or systemic levels of one or more cholesterol derivatives compared to baseline. In some embodiments, the one or more cholesterol derivatives are byproducts of cholesterol biosynthesis. In some embodiments, the one or more cholesterol derivatives include hydrogenation products, products containing 1H-cyclopenta[a]phenanthrene-3-ol products hydrogenated in different forms, or products formed having a hydroxyl group, an epoxy group, or a keto group. Optionally, the one or more cholesterol derivatives are oxysterols or sterols.

[0039] In some embodiments, the therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin is a suitable amount for achieving the therapeutic effect described herein. In some embodiments, the therapeutically effective amount is at least about 50 mg / kg, at least about 100 mg / kg, at least about 200 mg / kg, at least about 300 mg / kg, at least about 400 mg / kg, at least about 500 mg / kg, at least about 600 mg / kg, at least about 700 mg / kg, at least about 800 mg / kg, at least about 900 mg / kg, at least about 1000 mg / kg, at least about 1100 mg / kg, at least about 1200 mg / kg The amount is at least about 1300 mg / kg, at least about 1400 mg / kg, at least about 1500 mg / kg, at least about 1600 mg / kg, at least about 1700 mg / kg, at least about 1800 mg / kg, at least about 1900 mg / kg, at least about 2000 mg / kg, at least about 2100 mg / kg, at least about 2200 mg / kg, at least about 2300 mg / kg, at least about 2400 mg / kg, or at least about 2500 mg / kg. In some embodiments, the therapeutic effective dose of 2-hydroxypropyl-β-cyclodextrin is at least about 100 mg / kg. In some embodiments, the therapeutic effective dose of 2-hydroxypropyl-β-cyclodextrin is at least about 250 mg / kg. In some embodiments, the therapeutic effective dose of 2-hydroxypropyl-β-cyclodextrin is at least about 500 mg / kg. In some embodiments, the therapeutically effective dose of the 2-hydroxypropyl-β-cyclodextrin is at least about 1000 mg / kg. In some embodiments, the therapeutically effective dose of the 2-hydroxypropyl-β-cyclodextrin is at least about 1500 mg / kg.

[0040] In some embodiments, the therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin is a suitable amount for achieving the therapeutic effect described herein. In some embodiments, the therapeutically effective amount is about 50 mg / kg to about 2500 mg / kg (for example, about 50 mg / kg to about 1000 mg / kg, about 500 mg / kg to about 1000 mg / kg, about 500 mg / kg to about 1500 mg / kg, about 800 mg / kg to about 1500 mg / kg, about 800 mg / kg to about 1200 mg / kg, about 1000 mg / kg to about 1500 mg / kg, about 1000 mg / kg to about 2500 mg / kg). In some embodiments, the therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin is about 500 mg / kg to about 1500 mg / kg. In some embodiments, the therapeutically effective dose of the above 2-hydroxypropyl-β-cyclodextrin is approximately 800 mg / kg to approximately 1200 mg / kg.

[0041] In some embodiments, the therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin is an amount suitable for achieving the therapeutic effect described herein. In some embodiments, the therapeutically effective amount is at least about 4 g (for example, at least about 10 g, at least about 25 g, at least about 50 g, at least about 75 g, at least about 100 g, at least about 125 g, at least about 150 g, at least about 175 g, at least about 200 g, at least about 250 g). In some embodiments, the therapeutically effective dose of 2-hydroxypropyl-β-cyclodextrin may be about 4 g to about 250 g (e.g., about 4 g to about 200 g, about 4 g to about 150 g, about 4 g to about 100 g, about 4 g to about 50 g, about 50 g to about 250 g, about 50 g to about 200 g, about 50 g to about 150 g, about 50 g to about 100 g, about 100 g to about 250 g, about 100 g to about 200 g). The total amount of 2-hydroxypropyl-β-cyclodextrin administered (e.g., in a single dose, e.g., in a therapeutically effective dose) may depend on a number of factors, including, but not limited to, the age, sex, and weight of the subject.

[0042] In some embodiments, the therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin is sufficient to achieve whole blood, serum, and / or plasma concentrations of 2-hydroxypropyl-β-cyclodextrin suitable for achieving the therapeutic effects described herein. In some embodiments, the whole blood, serum, and / or plasma concentrations are at least about 0.01 mM (e.g., at least about 0.05 mM, at least about 0.1 mM, at least about 0.2 mM, at least about 0.3 mM, at least about 0.4 mM, at least about 0.5 mM, at least about 0.6 mM, at least about 0.7 mM, at least about 0.8 mM, at least about 0.9 mM, at least about 1.0 mM, at least about 1.5 mM, at least about 2.0 mM, at least about 2.5 mM, or at least about 3 mM).

[0043] The therapeutically effective dose may be the amount of 2-hydroxypropyl-β-cyclodextrin effective in raising the circulating and / or systemic levels of one or more oxysterols in the individual after administration compared to before administration. In some cases, the therapeutically effective dose is the amount of 2-hydroxypropyl-β-cyclodextrin effective in raising the circulating and / or systemic levels of one or more oxysterols in the individual (e.g., 1 hour after administration) by at least about 10%, for example, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% compared to before administration. In some embodiments, the one or more oxysterols are 24S-hydroxycholesterol, 27-hydroxycholesterol, or both.

[0044] The therapeutically effective dose may be the amount of 2-hydroxypropyl-β-cyclodextrin effective in increasing the cholesterol crystal solubility (CCDC) of plasma after administration (e.g., 1 hour after administration) compared to before administration. In some cases, the therapeutically effective dose is the amount of 2-hydroxypropyl-β-cyclodextrin effective in increasing plasma CCDC after administration (e.g., 1 hour after administration) by at least about 10%, for example, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% or more compared to before administration.

[0045] The therapeutically effective dose may be the amount of 2-hydroxypropyl-β-cyclodextrin effective in increasing the mRNA levels of genes regulated by one or more LXR transcription factors (e.g., ABCA1, ABCG1) after administration (e.g., 24 hours after administration) compared to before administration. In some cases, the therapeutically effective dose is the amount of 2-hydroxypropyl-β-cyclodextrin effective in increasing the mRNA levels of ABCA1 and / or ABCG1 after administration (e.g., 24 hours after administration) by at least about 10%, for example, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% or more compared to before administration.

[0046] The method disclosed herein may further comprise administering a first therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to a subject at a first time point, and administering a second therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the subject at a second time point. The second time point may be at least 4 hours, at least 6 hours, at least 8 hours, at least 12 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the first time point. In some embodiments, the administration may be by intravenous administration.

[0047] In some cases, the second timing may be determined based on one or more indicators that the additional administration of the drug is beneficial to the subject. For example, the second timing may be administered after the therapeutic benefit of the first administration has decreased or begun to decrease.

[0048] In various embodiments, the subjects described above may be human. In some cases, the subjects may be subjects of any age at which there is a risk of developing elevated levels of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) and / or developing CCE, or where such elevated levels of crystals (and / or blood clots) are more likely to occur and / or where CCE is more likely to occur. The subjects may be at least 30 years old (e.g., at least 40, at least 50, at least 60, at least 70, at least 80, at least 90). The subjects may have been diagnosed with atherosclerosis and / or atherosclerotic cardiovascular disease. In some cases, the subjects may have advanced atherosclerosis. In some cases, the subjects may have undergone vascular medical procedures such as vascular surgery or angiography. In some cases, the subjects may have initiated treatment with anticoagulants or thrombolytic agents. The subjects may have one or more risk factors for CCE. Risk factors for CCE include, but are not limited to, interventional vascular procedures, interventional diagnostic procedures, cardiovascular surgery, cardiovascular disease (e.g., coronary artery disease, atherosclerotic cardiovascular disease), aortic aneurysm, aortic plaque, hypertension, diabetes mellitus, hyperlipidemia, smoking, being male, age, increased inflammation (e.g., elevated serum (hs)CRP levels), anticoagulation therapy, and thrombolytic therapy.

[0049] The subjects described above may have been diagnosed with CCE and / or may have symptoms and / or clinical manifestations of CCE. CCE can be diagnosed, for example, by biopsy (e.g., skin biopsy, muscle biopsy, kidney biopsy, bone marrow biopsy, gastric mucosal biopsy, colonic mucosal biopsy). In some cases, the subjects can be diagnosed by a combination of disease-inducing events (e.g., cardiovascular surgery) and characteristic symptoms (e.g., symptoms of the skin, kidneys, central nervous system, and eyes as described herein (e.g., Hollenhorst spots)). In some cases, the subjects can be diagnosed by non-invasive imaging modalities (e.g., abdominal ultrasound, chest / abdominal computed tomography (CT), transthoracic echocardiography (TTE), transesophageal echocardiography (TEE)).

[0050] The above subjects may have one or more analytical test results consistent with CCE. Examples of one or more analytical test results consistent with CCE include, but are not limited to, elevated serum creatinine, leukocytosis, eosinophilia, anemia, thrombocytopenia, hypocomplementemia, elevated erythrocyte sedimentation rate, elevated (hs)CRP levels, elevated fibrinogen levels, eosinophilia, proteinuria, hematuria, and abnormal liver enzymes.

[0051] The subjects described above may be treated (for example, by the method described herein) before the onset of CCE (for example, after an inducing event such as cardiovascular surgery). For example, subjects at risk of developing CCE (for example, subjects with high levels of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals)) may be treated (for example, by the method described herein) to reduce the amount and / or size of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) and / or to change the shape of the crystals (and / or blood clots) (for example, thereby reducing the risk of developing CCE). In some cases, subjects with one or more CCE risk factors may be treated before an increase in levels of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) occurs. The subjects described above may be treated (for example, by the method described herein) after the onset of CCE and / or its symptoms or clinical manifestations.

[0052] The methods disclosed herein can be used to treat CCE and / or one or more symptoms and / or clinical symptoms of CCE. For example, the methods disclosed herein can be used to treat skin symptoms of CCE (e.g., reticular dermatosis, cyanosis, gangrene, skin ulcers, purpura, erythematous nodules, blue toe syndrome); atherothrombotic renal disease and / or renal symptoms of CCE (e.g., acute kidney injury, subacute kidney injury, chronic kidney injury, malignant hypertension, glomerulonephritis, end-stage renal disease, allograft dysfunction, renal infarction); gastrointestinal symptoms of CCE (e.g., abdominal pain, diarrhea, bleeding, intestinal ischemia, intestinal infarction, intestinal perforation, necrotizing pancreatitis, focal hepatocyte necrosis, acalculous cholecystitis); and central nervous system symptoms of CCE (e.g., headache, dizziness It can be used to treat dizziness, confusion, memory loss, transient ischemic attack, stroke, cerebral infarction, spinal cord infarction, paraplegia, mononeuropathy), ocular symptoms of CCE (e.g., transient monocular blindness, eye pain, blurred vision, Hollenhorst spots), myocardial infarction, adrenal insufficiency, penile necrosis, myositis, rhabdomyolysis, splenic infarction, alveolar hemorrhage, and / or symptoms associated with CCE (e.g., fever, fatigue, loss of appetite, weight loss, muscle pain).

[0053] In some embodiments, the methods described herein reduce the size (e.g., average size, maximum size) of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) in the subject. The size (e.g., average size, maximum size) of the circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) may be reduced compared to the size (e.g., average size, maximum size) of the circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) prior to the treatment. In some embodiments, the size (e.g., average size, maximum size) of the circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) may be reduced by at least about 0.5%. In some embodiments, the size (e.g., average size, maximum size) of the circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) may be reduced by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, and at least about 90% or more.

[0054] In some embodiments, the methods described herein reduce the amount (e.g., concentration) of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) in the subject. The amount (e.g., concentration) of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) may be reduced compared to the amount of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) prior to the treatment. In some embodiments, the amount (e.g., concentration) of circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) may be reduced by at least about 0.5%. In some embodiments, the amount (e.g., concentration) of the circulating (e.g., blood, plasma, serum) cholesterol crystals (and / or blood clots containing cholesterol crystals) may be reduced by at least about 0.5%, at least about 1%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, and at least about 90% or more. In some embodiments, the shape of the circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) is altered by the method described herein.

[0055] In some cases, the method described herein reduces inflammation (as determined, for example, by cytokine protein levels and / or RNA levels) compared to the level of inflammation prior to treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the method described herein improves renal function compared to the level of renal function prior to treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the method described herein improves dermatological symptoms compared to the level of dermatological symptoms prior to treatment with 2-hydroxypropyl-β-cyclodextrin. In some cases, the method described herein improves eosinophilia compared to the level of

[0056] In some embodiments, the above method includes treating a subject (e.g., having CCE, suspected of having CCE, or at risk of developing CCE) with a combination of 2-hydroxypropyl-β-cyclodextrin and an additional therapeutic agent.

[0057] Depending on the case, the additional therapeutic agents described above may be selected from the group consisting of HMG-CoA reductase inhibitors (statins), anti-inflammatory drugs (e.g., acetylsalicylic acid, colchicine, canakinumab), corticosteroids, immunosuppressants (e.g., cyclophosphamide), and proprotein convertase subtilisin / kexin type 9 (PCSK9) inhibitors.

[0058] Depending on the circumstances, 2-hydroxypropyl-β-cyclodextrin and the further therapeutic agents described above may be administered to the subject simultaneously or nearly simultaneously (e.g., as a single formulation or as separate formulations). Depending on the circumstances, 2-hydroxypropyl-β-cyclodextrin and the further therapeutic agents described above may be administered at different times (e.g., as separate formulations). Depending on the circumstances, the further therapeutic agent may be administered before the administration of 2-hydroxypropyl-β-cyclodextrin. Depending on the circumstances, the further therapeutic agent may be administered simultaneously with 2-hydroxypropyl-β-cyclodextrin. Depending on the circumstances, the further therapeutic agent may be administered after the administration of 2-hydroxypropyl-β-cyclodextrin.

[0059] In some cases, the subjects may have previously received treatment with additional therapeutic agents (for example, before administration of 2-hydroxypropyl-β-cyclodextrin). In some cases, such additional therapeutic agents may be ineffective or have limited efficacy. In such cases, subjects treated with 2-hydroxypropyl-β-cyclodextrin (for example, after or concurrently with such additional therapeutic agents) may show a greater therapeutic effect than those treated with such additional therapeutic agents alone.

[0060] In some cases, subjects treated with both 2-hydroxypropyl-β-cyclodextrin and additional therapeutic agents may exhibit a greater therapeutic effect than those shown by treatment with either the additional therapeutic agent or 2-hydroxypropyl-β-cyclodextrin alone. In some cases, treatment with both the additional therapeutic agent and 2-hydroxypropyl-β-cyclodextrin may have a synergistic effect, resulting in a greater overall effect from the interaction between the additional therapeutic agent and 2-hydroxypropyl-β-cyclodextrin than the sum of the individual effects of each agent. In some cases, treatment with both the additional therapeutic agent and 2-hydroxypropyl-β-cyclodextrin may have an additive effect.

[0061] Pharmaceutical composition

[0062] This specification discloses pharmaceutical compositions comprising, in certain embodiments, an amount of 2-hydroxypropyl-β-cyclodextrin effective for treating CCE and / or one or more symptoms and / or clinical manifestations of CCE in humans, and an excipient. The excipient may be a pharmaceutically acceptable excipient.

[0063] The above excipients may include tonicity modifiers, preservatives, solubilizers, buffers, solutions (e.g., solutions for intravenous administration), or any combination thereof. The tonicity modifier may be dextrose, glycerol, sodium chloride, glycerin, mannitol, or a combination thereof. The preservative may be an antioxidant, antibacterial agent, chelating agent, or a combination thereof. The antioxidant may be ascorbic acid, acetylcysteine, sulfites (e.g., bisulfite, metabisulfite), monothioglycerol, or a combination thereof. The antibacterial agent may be phenol, metacresol, benzyl alcohol, parabens, benzalkonium chloride, chlorobutanol, thimerosal, phenylmercury salts (e.g., acetate, borate, nitrate), or a combination thereof. The chelating agent may be ethylenediaminetetraacetic acid (EDTA) calcium disodium, EDTA disodium, EDTA sodium, calcium bercetamide sodium, cartelidol, diethylenetriaminepentaacetic acid (DTPA), or a combination thereof. The solubilizing agent may be a surfactant or a co-solvent. The surfactant may be polyoxyethylene sorbitan monooleate (Tween 80), sorbitan monooleate, polyoxyethylene sorbitan monolaurate (Tween 20), lecithin, polyoxyethylene-polyoxypropylene copolymer (Pluronics), or a combination thereof. The co-solvent may be propylene glycol, glycerin, ethanol, polyethylene glycol (PEG), sorbitol, dimethylacetamide, Cremophor EL, or a combination thereof. The polyethylene glycol may be PEG 300, PEG 400, PEG 600, PEG 3350, or PEG 4000.The above buffering agent may contain sodium acetate, acetic acid, glacial acetic acid, ammonium acetate, ammonium sulfate, ammonium hydroxide, arginine, aspartic acid, benzenesulfonic acid, sodium benzoate, benzoic acid, sodium bicarbonate, boric acid, sodium borate, sodium carbonate, citric acid, sodium citrate, disodium citrate, trisodium citrate, diethanolamine, glucono delta-lactone, glycine, glycine HCl, histidine, histidine HCl, hydrochloric acid, hydrobromic acid, lysine, maleic acid, meglumine, methanesulfonic acid, monoethanolamine, phosphate acid, monobasic potassium, dibasic potassium, monosodium phosphate, disodium phosphate, trisodium phosphate, sodium hydroxide, sodium succinate, sulfuric acid, sodium tartrate, tartaric acid, tromethamine (Tris), or a combination thereof.

[0064] The above pharmaceutical composition may contain at least about 4 g, at least about 10 g, at least about 50 g, at least about 100 g, at least about 150 g, at least about 200 g, or at least about 250 g of 2-hydroxypropyl-β-cyclodextrin. In some embodiments, the above pharmaceutical composition contains at least about 4 g of 2-hydroxypropyl-β-cyclodextrin. In some embodiments, the above pharmaceutical composition contains at least about 50 g of 2-hydroxypropyl-β-cyclodextrin. In some embodiments, the above pharmaceutical composition contains at least about 100 g of 2-hydroxypropyl-β-cyclodextrin. In some embodiments, the above pharmaceutical composition contains at least about 200 g of 2-hydroxypropyl-β-cyclodextrin. In some embodiments, the pharmaceutical composition contains about 4 g to about 250 g of 2-hydroxypropyl-β-cyclodextrin (for example, about 4 g to about 100 g, about 4 g to about 50 g, about 50 g to about 150 g, about 50 g to about 250 g, about 100 g to about 200 g, about 100 g to about 250 g, about 150 g to about 250 g).

[0065] The above-mentioned pharmaceutical composition may contain an amount of 2-hydroxypropyl-β-cyclodextrin effective in increasing the circulating and / or systemic levels of one or more oxysterols in the subject by at least about 10%, for example, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% or more after administration of the pharmaceutical composition to the subject (for example, 24 hours later).

[0066] The above-mentioned pharmaceutical composition may contain an amount of 2-hydroxypropyl-β-cyclodextrin that is effective in increasing the cholesterol crystal solubility (CCDC) of plasma in the subject by at least about 10%, for example, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% or more after administration of the pharmaceutical composition to the subject (for example, 1 hour later).

[0067] The above-mentioned pharmaceutical composition may contain an amount of 2-hydroxypropyl-β-cyclodextrin effective in increasing the mRNA level of a gene regulated by one or more LXR transcription factors (e.g., ABCA1 and / or ABCG1) by at least about 10%, for example, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50% or more after administration of the pharmaceutical composition to a subject (e.g., 24 hours later).

[0068] The above pharmaceutical composition may be formulated for single-dose administration. The above pharmaceutical composition may be formulated for intravenous administration. The above pharmaceutical composition may be formulated to be isotonic.

[0069] kit

[0070] Kits are further provided herein. Optionally, the kit comprises one or more containers (e.g., vials, flasks, wide-mouthed bottles, ampoules, etc.) containing one or more pharmaceutical compositions provided herein (e.g., 2-hydroxypropyl-β-cyclodextrin and pharmaceutically acceptable excipients). Optionally, the kit comprises multiple containers (e.g., two, three, four, five, six, seven, eight, nine, or ten or more containers). Optionally, at least one of the one or more containers is an intravenous infusion bag. The one or more containers may contain a single dose of the pharmaceutical composition, or multiple doses (e.g., two, three, four, five, six, seven, eight, nine, or ten or more doses). Optionally, the one or more containers may contain a high concentration of the pharmaceutical composition, which is then diluted before administration to obtain an effective dose. The dosage may be any amount as specified herein that is effective in treating one or more of the indications described herein. The kit may further comprise one or more further components for intravenous infusion of the pharmaceutical composition. Optionally, the kit may comprise an intravenous infusion bag. Optionally, the kit may comprise one or more solutions (e.g., physiological saline) for mixing and / or diluting the pharmaceutical composition. Optionally, the kit may comprise one or more of a catheter, tubing, syringe, and needle. The kit may further comprise instructions for administering the pharmaceutical composition for use in treating any of the indications described herein, for example, (e.g., to reduce the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in an individual, to reduce the size of the crystals and / or blood clots, and / or to alter the shape of the crystals and / or blood clots, and / or to treat cholesterol crystal embolism (CCE) and / or one or more of its symptoms in an individual). The kit may be supplied in a box, bag, or any other appropriate container.

[0071] In some embodiments, the kit may comprise one or more further active pharmaceutical ingredients (e.g., therapeutic compounds, drugs, etc.). In some cases, the kit may comprise a single container containing the pharmaceutical composition of the Disclosure (e.g., 2-hydroxypropyl-β-cyclodextrin and pharmaceutically acceptable excipients) and one or more further active pharmaceutical ingredients. In other cases, the kit may comprise a first container containing the pharmaceutical composition of the Disclosure (e.g., 2-hydroxypropyl-β-cyclodextrin and pharmaceutically acceptable excipients) and a second container containing one or more further active pharmaceutical ingredients. [Examples]

[0072] Example 1: Plasma obtained from subjects treated with 2-hydroxypropyl-β-cyclodextrin exhibits cholesterol crystal solubility. Male human subjects were treated with a single, escalating dose of 2-hydroxypropyl-β-cyclodextrin administered intravenously every four weeks according to Table 1 below. [Table 1]

[0073] For each administration, whole blood was collected before administration, after administration (line flush), 1 hour after administration, and 24 hours after administration. Plasma was separated from the whole blood and subjected to a cholesterol crystal lysis (CCDC) assay using a technique similar to that described in the literature. The CCDC assay measures the ability of the sample to dissolve cholesterol crystals.

[0074] Figure 1A shows the results of the CCDC assay described above. Figure 1A shows that the ability of plasma to dissolve cholesterol crystals increases after treatment with 2-hydroxypropyl-β-cyclodextrin. This suggests that treatment with 2-hydroxypropyl-β-cyclodextrin increases plasma factors involved in the dissolution of cholesterol crystals.

[0075] Plasma before administration was also incubated ex vivo with 2-hydroxypropyl-β-cyclodextrin, and its ability to dissolve cholesterol crystals was measured. Figure 1B shows that plasma treated with 2-hydroxypropyl-β-cyclodextrin ex vivo exhibits improved ability to dissolve cholesterol crystals.

[0076] In summary, the data presented herein demonstrate that, in some embodiments, treatment of humans with 2-hydroxypropyl-β-cyclodextrin can improve the cholesterol crystal solubility of plasma, thereby reducing the amount and / or size of circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) and / or altering the shape of the crystals (and / or blood clots). The data further demonstrate that, in some embodiments, treatment of humans with 2-hydroxypropyl-β-cyclodextrin, as described herein, may be a suitable treatment for cholesterol crystal embolism.

[0077] Example 2: Treatment with 2-hydroxypropyl-β-cyclodextrin increases sterol and oxysterol concentrations in human subjects. In this example, male human subjects were treated with 2-hydroxypropyl-β-cyclodextrin according to Example 1, and plasma levels of 24S-hydroxycholesterol and 27-hydroxycholesterol were measured. Figures 2A–2C show that treatment with 2-hydroxypropyl-β-cyclodextrin increased plasma levels of 24S-hydroxycholesterol and 27-hydroxycholesterol, while total cholesterol levels remained stable. 27-hydroxycholesterol is an endogenous LXR ligand that, in some embodiments, may induce downstream anti-inflammatory gene signatures that address the adverse inflammatory response mediated by the NLRP3 inflammasome effect in cholesterol crystal embolism. The elevated 27-hydroxycholesterol levels are also a cellular marker of increased macrophage activation and cholesterol crystal phagocytosis and clearance activity. Therefore, the above data indicates that, in some embodiments, treatment of humans with 2-hydroxypropyl-β-cyclodextrin may reduce the amount and / or size of the circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) and / or alter the shape of the crystals (and / or blood clots). The above data further indicates that, in some embodiments, treatment of humans with 2-hydroxypropyl-β-cyclodextrin, as described herein, may be a suitable treatment for cholesterol crystal embolism.

[0078] Example 3: Treatment with 2-hydroxypropyl-β-cyclodextrin increases genes regulated by the LXR transcription factor. In this example, male human subjects were treated with 2-hydroxypropyl-β-cyclodextrin according to Example 1, and mRNA levels of the genes ABCA1 and ABCG1, which are regulated by the LXR transcription factor, were measured. Figures 3A–3D show that treatment with 2-hydroxypropyl-β-cyclodextrin increased mRNA levels of ABCA1 and ABCG1. This data indicates an increase in both ABCA1 and ABCG1 cholesterol transporters in peripheral blood. ABCA1 and ABCG1 are important cholesterol transporters involved in cholesterol efflux from cells, loading of HDL particles for reverse cholesterol transport (RCT), and excretion via the hepatic pathway, potentially reducing cholesterol and cholesterol crystal embolisms released into vascular structures and stabilizing ruptured atherosclerotic plaques. Therefore, the above data indicates that, in some embodiments, treatment of humans with 2-hydroxypropyl-β-cyclodextrin may reduce the amount and / or size of the circulating cholesterol crystals (and / or blood clots containing cholesterol crystals) and / or alter the shape of the crystals (and / or blood clots). The above data further indicates that, in some embodiments, treatment of humans with 2-hydroxypropyl-β-cyclodextrin, as described herein, may be a suitable treatment for cholesterol crystal embolism.

[0079] While preferred embodiments of the present disclosure have been shown and described herein, it will be clear to those skilled in the art that such embodiments are provided merely as examples. Those skilled in the art will now be able to conceive of numerous variations, modifications, and substitutions without departing from the present disclosure. It is important to understand that various alternatives to the embodiments of the disclosure described herein can be adopted in the practice of the present disclosure. The appended claims define the scope of the present disclosure, and the methods and structures within these claims, as well as their equivalents, are intended to be covered thereby. The invention described in the original claims of this application is listed below. [1] A method for reducing the amount, size, and / or shape of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in an individual, the method comprising administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual to reduce the amount of circulating cholesterol crystals in the individual, reduce the size of the crystals, and / or change the shape of the crystals. [2] The method according to [1], wherein the size of the circulating cholesterol crystals and / or blood clot containing cholesterol crystals is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or more than 50% compared to the size of the circulating cholesterol crystals and / or blood clot containing cholesterol crystals before the administration of the 2-hydroxypropyl-β-cyclodextrin. [3] The method according to any one of the preceding numbers, wherein the size of the circulating cholesterol crystals and / or the blood clot containing the cholesterol crystals is the average size or the maximum size. [4] The method according to any one of the preceding numbers, wherein the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or more than 50% compared to the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals prior to the administration of 2-hydroxypropyl-β-cyclodextrin. [5] The method according to any one of the preceding numbers, wherein the amount of circulating cholesterol crystals is equal to the concentration of circulating cholesterol crystals. [6] The method according to any one of the preceding numbers, wherein the circulating cholesterol crystals are present in the blood, plasma, or serum. [7] The method according to any one of the preceding items, wherein the treatment reduces the incidence or severity of cholesterol crystal embolism (CCE) in the individual. [8] A method for treating cholesterol crystal embolism (CCE) and / or one or more symptoms thereof in an individual, comprising administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual, thereby treating the cholesterol crystal embolism (CCE) and / or one or more symptoms thereof in the individual. [9] The method according to [8], wherein the treatment comprises reducing the size of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in the individual.

[10] The method according to [9], wherein the size of the circulating cholesterol crystals and / or the blood clot containing the cholesterol crystals is the maximum size or the average size.

[11] The method according to any one of [8] to

[10] , wherein the treatment comprises reducing the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in the individual.

[12] The method according to

[11] , wherein the amount of the circulating cholesterol crystals and / or the blood clot containing cholesterol crystals is the concentration of the circulating cholesterol crystals.

[13] The method according to any one of [8] to

[12] , wherein the treatment involves altering the shape of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in the individual.

[14] The method according to any one of [8] to

[13] , wherein the treatment reduces inflammation in the individual.

[15] The method according to

[14] , wherein the inflammation is determined by cytokine protein levels and / or RNA levels.

[16] The method according to any one of [8] to

[15] , wherein the treatment described herein improves renal function in the said individual.

[17] The method according to any one of [8] to

[16] , wherein the treatment improves the dermatological symptoms in the individual.

[18] The method according to any one of [8] to

[17] , wherein the treatment described above improves eosinophilia in the individual.

[19] The method according to any one of [8] to

[18] , wherein the treatment described above improves a hematological abnormality in the individual.

[20] The method according to any one of [8] to

[19] , wherein the treatment improves the complement level in the individual.

[21] The method according to any one of [8] to

[20] , wherein the treatment described above improves proteinuria in the individual.

[22] The method according to any one of the preceding items, wherein the effective therapeutic dose is approximately 50 mg / kg to approximately 2,500 mg / kg.

[23] The method according to any one of the preceding items, wherein the effective therapeutic dose is approximately 4 g to approximately 250 g.

[24] The method according to any one of the preceding numbers, wherein the therapeutically effective dose is sufficient to achieve a serum, plasma, and / or whole blood concentration of 2-hydroxypropyl-β-cyclodextrin of about 0.01 mM to about 3 mM.

[25] The method according to any one of the preceding claims, wherein the therapeutically effective dose is an amount effective in increasing the circulating and / or systemic levels of one or more oxysterols in the individual by at least about 10% compared to before the administration.

[26] The method according to

[25] , wherein the one or more oxysterols are 24S-hydroxycholesterol, 27-hydroxycholesterol, or both.

[27] The method according to any one of the preceding numbers, wherein the therapeutically effective dose is an amount effective in increasing the cholesterol lysis capacity (CCDC) of plasma by at least about 10% after administration compared to before administration.

[28] The method according to any one of the preceding numbers, wherein the therapeutically effective dose is an amount effective in increasing the mRNA levels of ABCA1 and / or ABCG1 by at least about 10% after administration compared to before administration.

[29] The method according to any one of the preceding numbers, wherein the 2-hydroxypropyl-β-cyclodextrin is selected from the group consisting of Kleptose® HP parenteral grade, Kleptose® HPB parenteral grade, Kleptose® HPB-LB parenteral grade, Cavitron® W7 HP5 Pharma cyclodextrin, Cavitron® W7 HP7 Pharma cyclodextrin, Trappsol® Cyclo®, and VTS-270 / Adorabetadex.

[30] The method according to any one of the preceding items, wherein the individual has one or more CCE risk factors.

[31] The method according to

[30] , wherein one or more of the risk factors for CCE are selected from the group consisting of interventional vascular procedures, interventional diagnostic procedures, cardiovascular surgery, cardiovascular disease (e.g., coronary artery disease, atherosclerotic cardiovascular disease), aortic aneurysm, aortic plaque, hypertension, diabetes mellitus, hyperlipidemia, smoking, being male, age, increased inflammation (e.g., elevated serum (hs)CRP levels), anticoagulant therapy, thrombolytic therapy, and any combination thereof.

[32] The method according to any one of the preceding numbers, wherein the individual has obtained one or more analytical test results associated with CCE.

[33] The method according to

[32] , wherein one or more analytical test results associated with the CCE are selected from the group consisting of elevated serum creatinine, leukocytosis, eosinophilia, anemia, thrombocytopenia, hypocomplementemia, elevated erythrocyte sedimentation rate, elevated (hs)CRP levels, elevated fibrinogen levels, eosinophilia, proteinuria, hematuria, abnormal liver enzymes, and any combination thereof.

[34] The method according to any one of the preceding numbers, wherein the individual is at least 30 years old.

[35] The method according to any one of the preceding numbers, wherein the individual is a human.

[36] The aforementioned administration (i) At a first time point, administer to the individual a first dose of 2-hydroxypropyl-β-cyclodextrin that is therapeutically effective, (ii) At the second time point, administer to the individual a second dose of 2-hydroxypropyl-β-cyclodextrin that is therapeutically effective. The method described in any one of the preceding numbers, further including the method described in any one of the preceding numbers.

[37] The method according to

[36] , wherein the second time point is at least 4 hours, at least 6 hours, at least 8 hours, at least 12 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the first time point.

[38] The method described in any one of the preceding items, wherein the administration is by intravenous administration.

[39] An amount of 2-hydroxypropyl-β-cyclodextrin effective in reducing the quantity, size, and / or altering the shape of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in an individual, Pharmaceutically acceptable excipients and A pharmaceutical composition containing the following:

[40] A quantity of 2-hydroxypropyl-β-cyclodextrin effective in treating cholesterol crystal embolism (CCE) and / or its symptoms in an individual, Pharmaceutically acceptable excipients and A pharmaceutical composition containing the following:

[41] The pharmaceutical composition according to

[39] or 40, wherein the amount of 2-hydroxypropyl-β-cyclodextrin is an amount effective in increasing the circulating level and / or systemic level of one or more oxysterols in the individual by at least about 10% after administration of the pharmaceutical composition to the individual.

[42] The pharmaceutical composition according to

[41] , wherein the one or more oxysterols are 24S-hydroxycholesterol, 27-hydroxycholesterol, or both.

[43] The pharmaceutical composition according to any one of

[39] to

[42] , wherein the amount of 2-hydroxypropyl-β-cyclodextrin is an amount effective in increasing the cholesterol crystal solubility (CCDC) of plasma in the individual by at least about 10% after administration of the pharmaceutical composition to the individual.

[44] The pharmaceutical composition according to any one of

[39] to

[43] , wherein the amount of 2-hydroxypropyl-β-cyclodextrin is sufficient to increase the mRNA levels of ABCA1 and / or ABCG1 in the individual by at least about 10% after administration of the pharmaceutical composition to the individual.

[45] A pharmaceutical composition formulated for single-dose administration, as described in any one of items

[39] to

[44] .

[46] A pharmaceutical composition according to any one of the items

[39] to

[45] , formulated for intravenous administration.

[47] (a) One or more containers, (b) A pharmaceutical composition according to any one of the

[39] to

[46] stored in one or more containers and A kit that includes the following:

[48] (c) The kit according to

[47] further comprising instructions for use of the pharmaceutical composition for reducing the amount, size, and / or altering the shape of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in an individual, and / or treating cholesterol crystal embolism (CCE) and / or one or more of its symptoms in an individual.

[49] The kit according to

[47] or 48, wherein at least one of the one or more containers is an intravenous infusion bag.

[50] The kit according to any one of

[47] to

[49] , wherein the one or more containers include a single container containing the pharmaceutical composition and one or more further active pharmaceutical ingredients.

[51] The kit according to any one of

[47] to

[50] , wherein the one or more containers include a first container containing the pharmaceutical composition and a second container containing one or more further active pharmaceutical ingredients.

[52] The kit according to any one of the items

[47] to

[51] further comprises one or more further components selected from the group consisting of intravenous infusion bags, catheters, tubes, needles, syringes, solutions, and any combination thereof.

Claims

1. A pharmaceutical composition comprising 2-hydroxypropyl-β-cyclodextrin used in a method for reducing the amount, reducing the size, and / or changing the shape of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in an individual, comprising administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual, thereby reducing the amount of circulating cholesterol crystals in the individual, reducing the size of the crystals, and / or changing the shape of the crystals, wherein the therapeutically effective amount is about 500 mg / kg to about 1500 mg / kg, the individual is a human, and the administration is by intravenous administration.

2. a) The size of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or more than 50% compared to the size of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals before the administration of 2-hydroxypropyl-β-cyclodextrin, and / or b) The size of the circulating cholesterol crystals and / or the blood clot containing the cholesterol crystals is the average size or the maximum size, and / or c) The pharmaceutical composition according to claim 1, wherein the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals is reduced by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, or more than 50% compared to the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals before administration of 2-hydroxypropyl-β-cyclodextrin.

3. a) The amount of circulating cholesterol crystals is equal to the concentration of circulating cholesterol crystals, and / or b) The circulating cholesterol crystals are present in the blood, plasma, or serum, and / or c) The pharmaceutical composition according to claim 1 or 2, wherein the treatment reduces the incidence or severity of cholesterol crystal embolism (CCE) in the individual.

4. A pharmaceutical composition comprising 2-hydroxypropyl-β-cyclodextrin for use in a method of treating cholesterol crystal embolism (CCE) and / or one or more symptoms thereof in an individual, comprising administering a therapeutically effective amount of 2-hydroxypropyl-β-cyclodextrin to the individual, thereby treating the cholesterol crystal embolism (CCE) and / or one or more symptoms thereof in the individual, wherein the therapeutically effective amount is about 500 mg / kg to about 1500 mg / kg, the individual is a human, and the administration is by intravenous administration.

5. a) The treatment includes reducing the size of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals in the individual, wherein the size of the circulating cholesterol crystals and / or blood clots containing cholesterol crystals may be the maximum size or the average size, and / or b) The treatment includes reducing the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals in the individual, wherein the amount of circulating cholesterol crystals and / or blood clots containing cholesterol crystals may be the concentration of circulating cholesterol crystals, and / or c) The treatment includes altering the shape of the circulating cholesterol crystals and / or the blood clot containing cholesterol crystals in the individual, and / or d) The treatment comprises reducing inflammation in the individual, and the inflammation may be determined by cytokine protein levels and / or RNA levels, and / or e) The treatment includes improving renal function in the individual, and / or f) The treatment includes improving the dermatological symptoms in the individual, and / or g) The treatment includes improving eosinophilia in the individual, and / or h) The treatment includes improving a hematological abnormality in the individual, and / or i) The treatment includes improving complement levels in the individual, and / or j) The pharmaceutical composition according to claim 4, wherein the treatment described above includes improving proteinuria in the individual.

6. a) The effective therapeutic dose is approximately 4 g to approximately 250 g, and / or b) The pharmaceutical composition according to any one of claims 1 to 5, wherein the therapeutically effective amount is sufficient to achieve a serum, plasma, and / or whole blood concentration of 2-hydroxypropyl-β-cyclodextrin of about 0.01 mM to about 3 mM.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the therapeutically effective amount is an amount effective in raising the circulating level and / or systemic level of one or more oxysterols in the individual after administration by at least about 10% compared to before administration, and the one or more oxysterols may be 24S-hydroxycholesterol, 27-hydroxycholesterol, or both.

8. The pharmaceutical composition according to any one of claims 1 to 6, wherein the therapeutically effective amount is an amount effective in increasing the cholesterol crystal solubility (CCDC) of plasma by at least about 10% after administration compared to before administration.

9. The pharmaceutical composition according to any one of claims 1 to 6, wherein the therapeutically effective amount is an amount effective in increasing the mRNA levels of ABCA1 and / or ABCG1 by at least about 10% after administration compared to before administration.

10. a) The 2-hydroxypropyl-β-cyclodextrin is selected from the group consisting of Kleptose® HP parenteral grade, Kleptose® HPB parenteral grade, Kleptose® HPB-LB parenteral grade, Cavitron® W7 HP5 Pharma cyclodextrin, Cavitron® W7 HP7 Pharma cyclodextrin, Trappsol® Cyclo®, and VTS-270 / Adorabetadex, and / or b) The pharmaceutical composition according to any one of claims 1 to 9, wherein the individual has one or more risk factors for CCE, and the one or more risk factors for CCE may be selected from the group consisting of interventional vascular procedures, interventional diagnostic procedures, cardiovascular surgery, cardiovascular disease (e.g., coronary artery disease, atherosclerotic cardiovascular disease), aortic aneurysm, aortic plaque, hypertension, diabetes mellitus, hyperlipidemia, smoking, being male, age, increased inflammation (e.g., elevated serum (hs) CRP levels), anticoagulant therapy, thrombolytic therapy, and any combination thereof.

11. a) The individual has obtained one or more analytical test results associated with CCE, and the one or more analytical test results associated with CCE may be selected from the group consisting of elevated serum creatinine, leukocytosis, eosinophilia, anemia, thrombocytopenia, hypocomplementemia, elevated erythrocyte sedimentation rate, elevated (hs)CRP levels, elevated fibrinogen levels, eosinophilia, proteinuria, hematuria, abnormal liver enzymes, and / or, b) The individual is at least 30 years old, and / or c) The pharmaceutical composition according to any one of claims 1 to 10, wherein the individual is a human.

12. a) The administration of the above (i) At a first time point, administer to the individual a first dose of 2-hydroxypropyl-β-cyclodextrin that is therapeutically effective, (ii) At the second time point, administer to the individual a second dose of 2-hydroxypropyl-β-cyclodextrin that is therapeutically effective. The pharmaceutical composition according to any one of claims 1 to 11, further comprising, wherein the second time point may be at least 4 hours, at least 6 hours, at least 8 hours, at least 12 hours, at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks after the first time point.

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