# US 10,501,577 B2 · Poloxamer composition free of long circulating material and methods for production and uses thereof

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- Number: US 10,501,577 B2
- Document ID: US10501577B2
- Status: Granted
- Kind: B2
- Application number: 15/207,441
- Filed: 2016-07-11
- Published: 2017-01-05
- Granted: 2019-12-10
- Earliest claimed: 2014-07-07
- Assignees: LifeRaft Biosciences, Inc.
- Applicants: LifeRaft Biosciences, Inc.
- Inventors: R. Martin Emanuele; Mannarsamy Balasubramanian; Stewart V. Smith
- Web page: https://patentgrep.com/patent/US10501577B2

## CPC

- C08G 65/30: Post-polymerisation treatment, e.g. recovery, purification, drying
- A61K 9/0019: {Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00)}
- A61K 31/77: of oxiranes
- A61K 35/14: Blood; Artificial blood (perfluorocarbons A61K31/02; umbilical cord blood A61K35/51; haemoglobin A61K38/42)
- A61K 35/18: Erythrocytes (haemoglobin A61K38/42)
- A61K 35/19: Platelets; Megacaryocytes
- A61P 1/12: Antidiarrhoeals
- A61P 7/00: Drugs for disorders of the blood or the extracellular fluid
- A61P 7/02: Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
- A61P 7/04: Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
- A61P 9/00: Drugs for disorders of the cardiovascular system
- A61P 9/04: Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 25/00: Drugs for disorders of the nervous system
- A61P 27/02: Ophthalmic agents
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- C08G 65/08: Saturated oxiranes
- A61K 2035/124: {the cells being hematopoietic, bone marrow derived or blood cells}
- C08G 2650/58: Ethylene oxide or propylene oxide copolymers, e.g. pluronics
- C08G: MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Provided herein are long circulating material free (LCMF) poloxamer compositions and uses thereof. In particular, provided are LCMF poloxamer 188 compositions and uses thereof. Also provided are supercritical fluid extraction (SFE) methods and high pressure (subcritical) methods for preparing poloxamer compositions, particularly the LCMF poloxamer compositions.

## Claims

### Claim 1

A long circulating material free (LCMF) poloxamer 188, wherein:
the LCMF poloxamer 188 is a polyoxyethylene/polyoxypropylene copolymer that has the formula HO(CH2CH2O)a′—[CH(CH3)CH2O]b—(CH2CH2O)aH, wherein:
each of a and a′ is an integer such that the percentage of the hydrophile (C2H4O) is between approximately 60% and 90% by weight of the total molecular weight of the copolymer;
a and a′ are the same or different;
b is an integer such that the molecular weight of the hydrophobe (C3H6O) is between approximately 1,300 and 2,300 Daltons;
no more than 1.5% of the total components in the polymeric distribution of the co-polymer are low molecular weight components having an average molecular weight of less than 4,500 Daltons;
no more than 1.5% of the total components in the polymeric distribution of the co-polymer are high molecular weight components having an average molecular weight of greater than 13,000 Daltons;
the polydispersity value of the copolymer is less than approximately 1.07 or less than 1.07; and
following intravenous administration to a human subject, the circulating plasma half-life of any components not comprising the main peak in the distribution of copolymer is no more than 5.0-fold the circulating half-life of the main component in the distribution of the copolymer.

### Claim 2 (depends on 1)

The LCMF poloxamer of claim 1, wherein all components comprising the polymeric distribution of the copolymer have a circulating half-life in the plasma of the subject that is no more than 4.0-fold, or 3.0-fold longer than the circulating half-life of the main component of the co-polymer following intravenous administration to a subject.

### Claim 3 (depends on 1)

The LCMF poloxamer of claim 1, wherein all components in the distribution of the copolymer, when administered to a human subject, have a circulating half-life in the plasma of the subject that is no more than 3-fold longer than the circulating half-life of the main component in the distribution of the co-polymer.

### Claim 4 (depends on 1)

The LCMF poloxamer of claim 1, wherein all components in the distribution of the copolymer, when administered to a human subject, have a half-life in the plasma of the subject that is no more than 10 or 12 hours.

### Claim 5 (depends on 1)

The LCMF poloxamer of claim 1, wherein the polyoxyethylene/polyoxypropylene copolymer is a poloxamer with a hydrophobe having a molecular weight of about 1,400 to 2,000 Da or 1,400 to 2,000 Da, and a hydrophile portion constituting approximately 70% to 90% or 70% to 90% by weight of the copolymer.

### Claim 6 (depends on 1)

The LCMF poloxamer of claim 1, wherein: the molecular weight of the hydrophobe (C3H6O) is about or is 1,750 Da; and the average molecular weight of the polyoxyethylene/polyoxypropylene copolymer is 7,680 to 9,510 Daltons.

### Claim 7 (depends on 6)

The LCMF poloxamer of claim 6, wherein the average molecular weight of the polyoxyethylene/polyoxypropylene copolymer is 8,400-8,800 Daltons.

### Claim 8 (depends on 1)

The LCMF poloxamer 188 of claim 1, wherein:
the LCMF poloxamer 188 is a polyoxyethylene/polyoxypropylene copolymer that has the formula HO(CH2CH2O)a′—[CH(CH3)CH2O]b—(CH2CH2O)aH;
each of a and a′ is an integer such that the percentage of the hydrophile (C2H4O) is between approximately 60% and 90% by weight of the total molecular weight of the copolymer;
a and a′ are the same or different;
b is an integer such that the molecular weight of the hydrophobe (C3H6O) is between approximately 1,300 and 2,300 Daltons;
no more than 1.5% of the total components in the distribution of the co-polymer are low molecular weight components having an average molecular weight of less than 4,500 Daltons;
no more than 1.5% of the total components in the distribution of the co-polymer are high molecular weight components having an average molecular weight of greater than 13,000 Daltons;
the polydispersity value of the copolymer is less than approximately 1.07 or less than 1.07; and
the LCMF poloxamer is more hydrophilic than a purified poloxamer 188 that contains the long circulating material (LCM).

### Claim 9 (depends on 1)

The LCMF poloxamer 188 of claim 1, wherein:
the LCMF poloxamer 188 is a polyoxyethylene/polyoxypropylene copolymer that has the formula —HO(CH2CH2O)a′—[CH(CH3)CH2O]b—(CH2CH2O)aH—;
each of a and a′ is an integer such that the percentage of the hydrophile (C2H4O) is between approximately 60% and 90% by weight of the total molecular weight of the copolymer;
a and a′ are the same or different;
b is an integer such that the molecular weight of the hydrophobe (C3H6O) is between approximately 1,300 and 2,300 Daltons;
no more than 1.5% of the total components in the distribution of the co-polymer are low molecular weight components having an average molecular weight of less than 4,500 Daltons;
no more than 1.5% of the total components in the distribution of the co-polymer are high molecular weight components having an average molecular weight of greater than 13,000 Daltons;
the polydispersity value of the copolymer is less than approximately 1.07 or less than 1.07;
the LCMF poloxamer 188 has a mean retention time (tR) as assessed by reverse phase-high performance liquid chromatography (RP-HPLC) that is shorter than purified LCM-containing poloxamer 188 under the same RP-HPLC conditions; and
the capacity factor (k′) of LCMF poloxamer 188 as assessed by RP-HPLC is less than the k′ for purified LCM-containing poloxamer 188 under the same RP-HPLC conditions.

### Claim 10 (depends on 1)

The LCMF poloxamer of claim 1, produced by a method comprising supercritical fluid extraction pressurized with CO2.

### Claim 11 (depends on 1)

The LCMF poloxamer of claim 1, produced by a method comprising:
a) introducing a poloxamer 188 solution into an extractor vessel, wherein the poloxamer 188 is dissolved in a first alkanol to form a poloxamer 188 solution;
b) admixing the poloxamer 188 solution with an extraction solvent comprising a second alkanol and supercritical carbon dioxide under a temperature and pressure to maintain the supercritical carbon dioxide for a first defined period, wherein:
the temperature is above the critical temperature of carbon dioxide but is no more than 40° C.;
the pressure is 220 bars to 280 bars; and
the alkanol is provided at an alkanol concentration that is 7% to 8% by weight of the total extraction solvent; and
c) increasing the concentration of the second alkanol in step b) in the extraction solvent a plurality of times in gradient steps over time of the extraction method, wherein:
each plurality of times occurs for a further defined period; and in each successive step, the alkanol concentration is increased 1-2% compared to the previous concentration of the second alkanol; and
d) removing the extraction solvent from the extractor vessel to thereby remove the extracted material from the raffinate poloxamer preparation.

### Claim 12 (depends on 11)

The LCMF poloxamer of claim 11, wherein:
in step a), the ratio of poloxamer to first alkanol, by weight is about or is from 2:1 to 3:1, inclusive; and
the plurality of times in step c) occurs in two, three, four or five gradient steps.

### Claim 13 (depends on 1)

A pharmaceutical composition, comprising the LCMF poloxamer 188 of claim 1 in a pharmaceutically acceptable formulation.

### Claim 14 (depends on 13)

A method of treating a disease or condition in a subject, comprising administering the pharmaceutical composition of claim 13 to the subject, wherein the disease or condition is selected from the group consisting of heart failure, myocardial infarction, limb ischemia, shock, stroke, ischemic stroke, sickle cell disease, neurodegenerative diseases, macular degeneration, diabetic retinopathy and congestive heart failure.

### Claim 15 (depends on 14)

The method of claim 14, wherein the condition is heart failure that is acute heart failure or chronic heart failure.

### Claim 16 (depends on 14)

The method of claim 14, wherein the condition is acute myocardial infarction, acute limb ischemia or acute stroke.

### Claim 17 (depends on 14)

The method of claim 14, wherein the condition is sickle cell disease.

### Claim 18 (depends on 1)

A composition, comprising the long circulating material free (LCMF) poloxamer 188 of claim 1, wherein:
the composition is formulated for intravenous administration; and
the composition comprises 5-50 gm of the LCMF poloxamer.

### Claim 19 (depends on 1)

A composition, comprising an the LCMF poloxamer of claim 1, wherein:
the composition is formulated for intravenous administration; and
the composition comprises 5-50 gm of the LCMF poloxamer.

### Claim 20 (depends on 1)

A composition comprising the LCMF poloxamer 188 of claim 1 and one or more components selected from the group consisting of blood, and blood product.

### Claim 21 (depends on 20)

The composition of claim 20, wherein said one or more components is packed red blood cells or platelets.

### Claim 22 (depends on 21)

A transfusion method comprising administering the composition of claim 21 to a subject, wherein:
the subject has a disease or disorder or condition selected from the group consisting of sickle cell disease, acute chest syndrome, peripheral artery disease, heart failure, stroke, peripheral vascular disease, macular degeneration, respiratory distress syndrome (ARDS), multiple organ failure, ischemia, shock, acidosis, hypothermia, anemia, trauma, blood loss and blood disorders.

### Claim 23 (depends on 22)

The method of claim 22, wherein the disease, disorder or condition is selected from among hemorrhagic shock, septic shock, acute ARDS, anemic decompensation and blood loss.

### Claim 24 (depends on 1)

A method of preparing the long circulating material free (LCMF) poloxamer 188 of claim 1, comprising:
a) introducing a poloxamer 188 solution into an extractor vessel, wherein the poloxamer 188 is dissolved in a first alkanol to form a poloxamer 188 solution;
b) admixing the poloxamer 188 solution with an extraction solvent comprising a second alkanol and supercritical carbon dioxide under a temperature and pressure to maintain the supercritical carbon dioxide for a first defined period, wherein:
the temperature is above the critical temperature of carbon dioxide but is no more than 40° C.;
the pressure is 220 bars to 280 bars; and
the alkanol is provided at an alkanol concentration that is 7% to 8% by weight of the total extraction solvent;
c) increasing the concentration of the second alkanol in step b) in the extraction solvent a plurality of times in gradient steps over time of the extraction method, wherein:
each plurality of times occurs for a further defined period; and
in each successive step, the alkanol concentration is increased 1-2% compared to the previous concentration of the second alkanol; and
d) removing the extraction solvent from the extractor vessel to thereby remove the extracted material from the raffinate poloxamer preparation to thereby produce the LCMF poloxamer.

### Claim 25 (depends on 24)

The method of claim 24, wherein in step a), the ratio of poloxamer to first alkanol, by weight, is about or is from 2:1 to 3:1, inclusive.

### Claim 26 (depends on 24)

The method claim 24, wherein step c) occurs in two steps comprising:
i) increasing the concentration of the second alkanol from about 7% to 8% to about 8.1% to 9.5% for a second defined period; and
ii) increasing the concentration of the second alkanol from about 8.2% to 9.5% to about 9.6% to 11.5% for a third defined period.

### Claim 27 (depends on 24)

The method of claim 24, wherein the first defined period, second defined period and third defined period each are performed for 2 hours to 12 hours.

### Claim 28 (depends on 24)

The method of claim 24, wherein: the first and second alkanol are each independently selected from the group consisting of methanol, ethanol, propanol, butanol, pentanol and a combination thereof.

### Claim 29 (depends on 24)

The method of claim 24, wherein step d) of removing the extracted material occurs throughout steps b) and c).