# US 11,896,606 B2 · Pharmaceutical composition comprising particles and methods of making same

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- Number: US 11,896,606 B2
- Document ID: US11896606B2
- Status: Granted
- Kind: B2
- Application number: 17/095,059
- Filed: 2020-11-11
- Published: 2021-03-04
- Granted: 2024-02-13
- Earliest claimed: 2017-05-17
- Assignees: Highlight Therapeutics, S.L.
- Applicants: Highlight Therapeutics, S.L.
- Inventors: Mercedes Pozuelo Rubio; Marisol Quintero Ortiz; Ana Villanueva Garcia
- Web page: https://patentgrep.com/patent/US11896606B2

## CPC

- A61K 31/713: Double-stranded nucleic acids or oligonucleotides
- 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 9/08: Solutions {(composition of solutions A61K47/00)}
- A61K 9/19: lyophilised {, i.e. freeze-dried, solutions or dispersions (lyophilised products with subsequent particle size reduction A61K9/14; granules or pellets made by lyphilisation A61K9/1682; solid oral dosage forms made by lyophilisation A61K9/2095; lyophilisation additives A61K47/00)}
- A61K 31/00: Medicinal preparations containing organic active ingredients
- A61K 31/15: Oximes (>C=N—O—); Hydrazines (>N—N<); Hydrazones (>N—N=) {; Imines (C—N=C)}
- A61K 31/74: Synthetic polymeric materials
- A61K 39/3955: {against proteinaceous materials, e.g. enzymes, hormones, lymphokines}
- A61K 45/06: Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
- A61K 47/26: Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
- A61K 47/50: the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K 47/59: obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
- A61K 47/6929: {the form being a nanoparticle, e.g. an immuno-nanoparticle}
- A61K 47/6935: {the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol}
- A61P 31/04: Antibacterial agents
- A61P 31/12: Antivirals
- A61P 35/00: Antineoplastic agents
- C07K 16/2818: {against CD28 or CD152}
- C07K 16/2827: {against B7 molecules, e.g. CD80, CD86}
- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- 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
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

The present invention relates to compositions comprising particles, each of the respective particles comprising particles formed by polyribonucleotides and polymers. Specifically, the composition comprises a complex of at least one double-stranded polyribonucleotide, such as polyinosinic-polycytidylic acid [poly(I:C)], and at least one linear polyalkyleneimine. The particles are also characterized by their monomodal diameter distribution and z-average diameter within specific ranges. The present invention additionally relates to methods of manufacturing the compositions, and further to related pharmaceutical compositions useful for the treatment of any cell growth disorder characterized by an abnormal growth of human or animal cells.

## Claims

### Claim 1

An aqueous composition comprising one or more particles, wherein:
(a) each particle comprises a complex of at least one double-stranded polyribonucleotide, or a salt or solvate thereof, and at least one linear polyalkyleneimine, or a salt and/or solvate thereof, wherein
(i) the double-stranded polyribonucleotide is polyinosinic-polycytidylic acid [poly(I:C)], and
(ii) the weight average molecular weight of the linear polyalkyleneimine is between 17 and 23 kDa and has a molecular weight polydispersity index of below 1.5;
(b) at least 90% of the particles have a mono-modal diameter distribution of below 300 nm;
(c) the one or more particles each have a z-average diameter of less than or equal to 150 nm measured according to ISO 22412:2008; and
(d) the aqueous composition has a zeta potential equal to or greater than 30 mV measured according to ISO 13099-2:2012.

### Claim 2 (depends on 1)

The aqueous composition according to claim 1, wherein at least 90% of the particles have a mono-modal diameter distribution of between 30 nm and 150 nm.

### Claim 3 (depends on 1)

The aqueous composition according to claim 1, wherein at least 40% of the double-stranded polyribonucleotides comprised in the particles have at least 850 base pairs, and at least 50% of the double-stranded polyribonucleotides comprised in the particles have between 400 and 5000 base pairs.

### Claim 4 (depends on 1)

The aqueous composition according to claim 1, wherein the aqueous composition has a zeta potential of between 35 mV and 50 mV.

### Claim 5 (depends on 1)

The aqueous composition according to claim 1, wherein:
(i) between 5% and 60% of the double-stranded polyribonucleotides have less than 400 base pairs;
(ii) between 20% and 45% of the double-stranded polyribonucleotides have between 400 and 850 base pairs;
(iii) between 20% and 70% of the double-stranded polyribonucleotides have between 850 and 5000 base pairs; and/or
(iv) between 0% and 10% of the double-stranded polyribonucleotides have more than 5000 base pairs.

### Claim 6 (depends on 1)

The aqueous composition according to claim 1, wherein the linear polyalkyleneimine is a linear polyethyleneimine.

### Claim 7 (depends on 1)

The aqueous composition according to claim 1, wherein the weight average molecular weight and the polydispersity index of the linear polyalkyleneimine is determined for the polyalkyleneoxide precursor to the polyalkyleneimine as measured according to ISO 16014:2012.

### Claim 8 (depends on 1)

The aqueous composition according to claim 1, wherein the at least one double-stranded polyribonucleotide composition contains poly(I:C) at a concentration of at least 0.5 mg/ml.

### Claim 9 (depends on 1)

The aqueous composition according to claim 1, further comprising at least one pharmaceutically acceptable carrier, organic solvent, excipient and/or adjuvant.

### Claim 10 (depends on 1)

The aqueous composition according to claim 1, further comprising at least one compound selected from an organic compound, an inorganic compound, a nucleic acid, an aptamer, a peptide, and/or a protein.

### Claim 11 (depends on 1)

The aqueous composition according to claim 1, further comprising glucose or mannitol in a concentration of between 1 and 10% (weight/volume).

### Claim 12 (depends on 1)

The aqueous composition according to claim 1, wherein the aqueous composition has a pH of between 2 and 4, and/or an osmolarity of between 200 and 600 mOsm/kg.

### Claim 13 (depends on 1)

An aqueous composition comprising the one or more particles according to claim 1, wherein each respective particle is formed at a ratio of a number of moles of nitrogen of the polyalkyleneimine to a number of moles of phosphorus of the double-stranded polyribonucleotide that is equal to or greater than 2.5.

### Claim 14 (depends on 13)

The aqueous composition according to claim 13, wherein the aqueous composition is formed by a complex of at least 0.5 mg of poly(I:C) per ml of the total volume of the composition.

### Claim 15 (depends on 13)

An aqueous composition according to claim 13, wherein the aqueous composition is formed by additionally adding glucose or mannitol at a concentration of between 1 and 10% (weight/total volume of the composition).

### Claim 16 (depends on 1)

A composition obtained by lyophilizing the aqueous composition according to claim 1.

### Claim 17 (depends on 1)

A method for preparing an aqueous composition comprising the one or more particles of claim 1, comprising:
(a) preparing a first aqueous solution of at least one double-stranded polyribonucleotide, or a salt or solvate thereof, and a second aqueous solution of at least one polyalkyleneimine, or a salt or solvate thereof;
(b) independently filtering each respective first and second aqueous solution through a filter having a pore diameter of less than or equal to 500 nm thereby forming a corresponding first and second sterilized solution;
(c) mixing each resulting first and second sterilized solution in a mixing chamber thereby forming the aqueous composition,
(i) wherein one of the respective first and second sterilized solutions is introduced into the other respective sterilized solution in the mixing chamber, optionally by injection, at a rate of greater than or equal to 1 ml/min, or
(ii) wherein each respective first and second sterilized solution is introduced into the mixing chamber simultaneously, optionally by injection, at a rate of greater than or equal to 1 ml/min.

### Claim 18 (depends on 17)

The method according to claim 17, further comprising:
(d) filtering the resulting aqueous composition through a filter having a pore diameter of less than or equal to 600 nm to thereby form a filtrate, or centrifuging the resulting aqueous composition at a speed of greater than or equal to 22480 m/s2 to form a supernatant, wherein the resulting aqueous composition, filtrate or supernatant are optionally lypholized.

### Claim 19 (depends on 17)

The method according to claim 17, wherein one or both of the respective first and second aqueous solutions each further comprise a pharmaceutically acceptable carrier, organic solvent, excipient and/or adjuvant.