# US 2020/0399333 A1 · Histatins and method of use thereof

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- Number: US 2020/0399333 A1
- Document ID: US20200399333A1
- Status: Published application
- Kind: A1
- Application number: 17/011,273
- Filed: 2020-09-03
- Published: 2020-12-24
- Earliest claimed: 2015-11-30
- Assignees: The Board of Trustees of the University of Illinois
- Applicants: The Board of Trustees of the University of Illinois
- Inventors: Vinay AAKALU
- Web page: https://patentgrep.com/patent/US20200399333A1

## CPC

- C07K 14/4723: {Cationic antimicrobial peptides, e.g. defensins}
- A61K 38/00: Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00)
- A61K 38/1709: {from mammals}
- A61P 27/02: Ophthalmic agents
- A61K 9/0048: {Eye, e.g. artificial tears}
- A61K 9/0051: {Ocular inserts or implants}
- A61K 38/06: Tripeptides
- A61K 38/08: Peptides having 5 to 11 amino acids {(A61K38/043 - A61K38/046 take precedence)}
- A61K 38/12: Cyclic peptides {, e.g. bacitracins; Polymyxins; Gramicidins S, C; Tyrocidins A, B or C (A61K38/043 - A61K38/046 take precedence)}
- C12N 9/50: Proteinases {, e.g. Endopeptidases (3.4.21-3.4.25)}
- 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)
- 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
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Synthetic histatins composed of combinations of functional domains of natural histatins separated by exogenous linkers are described as are methods of using endogenous and synthetic histatins for the treatment of ocular diseases or conditions.

## Claims

### Claim 1

A method of treating an ocular disease or condition comprising administering to a subject in need of treatment an effective amount of a synthetic histatin having the general structure:
[HTNF1-L1-HTNF2-(L2)y]x (Formula I)
wherein
i) HTNF1 is a first histatin fragment ranging in length from 5 to 20 amino acids;
ii) HTNF2 is a second histatin fragment ranging in length from 5 to 20 amino acids;
iii) L1 is a first linker;
iv) L2 is a second linker;
v) x=1 to 3; and
vi) y=0 to 2;
wherein HTNF1 and HTNF2 are each independently the same or different and L1 and L2 are each independently the same or different, thereby treating the subject's ocular disease or condition.

### Claim 2 (depends on 1)

The method of claim 1, wherein HTNF1 or HTNF2 has an amino acid sequence of SNYLYDN (SEQ ID NO:1) or HEXXH (SEQ ID NO:2), wherein each X is independently a basic amino acid residue.

### Claim 3 (depends on 1)

The method of claim 1, wherein L1 or L2 is a flexible linker, rigid linker, in vivo cleavable linker, or a combination thereof.

### Claim 4 (depends on 3)

The method of claim 3, wherein the flexible linker is a hydrocarbon linker.

### Claim 5 (depends on 4)

The method of claim 4, wherein the hydrocarbon linker has the structure —(CH2)6—.

### Claim 6 (depends on 3)

The method of claim 3, wherein the flexible linker is a peptide linker having an amino acid sequence of (GGGGS)n (SEQ ID NO:3), KESGSVSSEQLAQFRSLD (SEQ ID NO:4), or EGKSSGSGSESKST (SEQ ID NO:5), GGGGGGGG (SEQ ID NO:6), GSAGSAAGSGEF (SEQ ID NO:7), (GGSG)n (SEQ ID NO:8), or (GS)n (SEQ ID NO:9), wherein n is 1, 2, 3, 4 or 5.

### Claim 7 (depends on 3)

The method of claim 3, wherein the rigid linker is a peptide linker having an amino acid sequence of (EAAAK)n (SEQ ID NO:10), A(EAAAK)nA (SEQ ID NO:11), PAPAP (SEQ ID NO:12), or (XP)n (SEQ ID NO:13), wherein n is 1, 2, 3, 4 or 5.

### Claim 8 (depends on 3)

The method of claim 3, wherein the in vivo cleavable linker is a peptide linker having an amino acid sequence of VSQTSKLTRAETVFPDV (SEQ ID NO:14), PLGLWA (SEQ ID NO:15), RVLAEA (SEQ ID NO:16), EDVVCCSMSY (SEQ ID NO:17), GGIEGRGS (SEQ ID NO:18), TRHRQPRGWE (SEQ ID NO:19), AGNRVRRSVG (SEQ ID NO:20), RRRRRRRRR (SEQ ID NO:21), GFLG (SEQ ID NO:22), or CRRRRRREAEAC (SEQ ID NO:23).

### Claim 9 (depends on 1)

The method of claim 1, wherein said synthetic histatin is linear or cyclized.

### Claim 10 (depends on 9)

The method of claim 9, wherein the synthetic histatin is cyclized via a disulfide bridge between terminal cysteine residues.

### Claim 11 (depends on 9)

The method of claim 9, wherein the synthetic histatin is cyclized with a sortase or butelase.

### Claim 12 (depends on 1)

The method of claim 1, wherein said synthetic histatin has the structure:
(a) GYKRKFHEKHHSHR(SEQ ID NO:24)-L1-YGDYGSNYLYDN(SEQ ID NO:25);
(b) HEKHH(SEQ ID NO:26)-L1-HEKHH(SEQ ID NO:26)-L2-HEKHH(SEQ ID NO:26)-L1-YGDYGSNYLYDN(SEQ ID NO:25);
(c) HEKRHH(SEQ ID NO:27)-L1-HEKRHH(SEQ ID NO:27)-L2-HEKRHH(SEQ ID NO:27)-L1-YGDYGSNYLYDN(SEQ ID NO:25);
(d) HEKRHH(SEQ ID NO:27)-L1-HEKRHH(SEQ ID NO:27)-L2-HEKHH(SEQ ID NO:26)-L1-YGDYGSNYLYDN(SEQ ID NO:25); or
(e) HEKRHH(SEQ ID NO:27)-L1-HEKHH(SEQ ID NO:26)-L2-HEKHH(SEQ ID NO:26)-L1-YGDYGSNYLYDN(SEQ ID NO:25),
wherein each L1 and L2 is independently a flexible linker, rigid linker, or in vivo cleavable linker.

### Claim 13 (depends on 1)

The method of claim 1, wherein said synthetic histatin has the structure:
(a)
(SEQ ID NO: 28)
GYKRKFHEKHHSHR-(CH2)6-YGDYGSNYLYDN;
(b)
(SEQ ID NO: 29)
HEKHH-(CH2)6-HEKHH-(CH2)6-HEKHH-(CH2)6-
YGDYGSNYLYDN;
(c)
(SEQ ID NO: 30)
HEKRHH-(CH2)6-HEKRHH-(CH2)6-HEKRHH-(CH2)6-
YGDYGSNYLYDN;
(d)
(SEQ ID NO: 31)
HEKRHH-(CH2)6-HEKRHH-(CH2)6-HEKHH-(CH2)6-
YGDYGSNYLYDN;
or
(e)
(SEQ ID NO: 32)
HEKRHH-(CH2)6-HEKHH-(CH2)6-HEKHH-(CH2)6-
YGDYGSNYLYDN

### Claim 14 (depends on 1)

The method of claim 1, wherein the ocular disease or condition is an ocular surface inflammatory disorder, dry eye disease, ocular tissue neovascularization, age-related macular degeneration, diabetic retinopathy, chronic or acute severe uveitis, a retinal pigment epithelial disorder, an ocular surface disease, an infection, or corneal or conjunctival wound.

### Claim 15 (depends on 1)

The method of claim 1, wherein the synthetic histatin provides wound healing, antimicrobial, metal ion chelating, anti-inflammatory, anti-angiogenic, or matrix metalloproteinase inhibitory activity.