# US 10,987,388 B2 · Homogeneous engineered phage populations

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,987,388 B2
- Document ID: US10987388B2
- Status: Granted
- Kind: B2
- Application number: 16/040,793
- Filed: 2018-07-20
- Published: 2019-01-24
- Granted: 2021-04-27
- Earliest claimed: 2017-07-21
- Assignees: Massachusetts Institute of Technology
- Applicants: Massachusetts Institute of Technology
- Inventors: Angela M. Belcher; Uyanga Tsedev; Fred Lam
- Web page: https://patentgrep.com/patent/US10987388B2

## CPC

- A61K 35/76: Viruses; Subviral particles; Bacteriophages
- A61K 47/6901: {Conjugates being cells, cell fragments, viruses, ghosts, red blood cells or viral vectors}
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- C12N 15/70: Vectors or expression systems specially adapted for E. coli
- C12N 15/861: Adenoviral vectors
- 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 47/6415: {Toxins or lectins, e.g. clostridial toxins or Pseudomonas exotoxins}
- A61K 49/0021: {the fluorescent group being a small organic molecule}
- C12N 2795/14121: Viruses as such, e.g. new isolates, mutants or their genomic sequences
- C12N 2795/14122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2795/14131: Uses of virus other than therapeutic or vaccine, e.g. disinfectant
- C12N 2795/14132: Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
- C12N 2800/202: of bacteriophage origin
- C12N 2810/40: Vectors comprising a peptide as targeting moiety, e.g. a synthetic peptide, from undefined source
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided are engineered phages populations, which are homogeneous in length, as well as methods of making and methods of using such phages. Also provided are engineered chlorotoxin-phages as well as their methods of making and using. The disclosed homogeneous phage populations and chlorotoxin-phages may be used, for example, for treating and/or imaging tumors, such as central nervous system tumors.

## Claims

### Claim 1

A homogeneous engineered filamentous phage population, wherein at least 70% of filamentous phages in the phage population have a length within 15% or within 8 nm of a length value, which is selected from 10 nm to less than the corresponding wild type filamentous phage length, wherein the phage population has a count of at least 1e13 pfu.

### Claim 2 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, wherein at least 70% of filamentous phages in the phage population have a length within 15% of the length value.

### Claim 3 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, wherein at least 90% of filamentous phages in the phage population have a length within 15% of the length value.

### Claim 4 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, wherein at least 70% of filamentous phages in the phage population have a length within 8 nm of the length value.

### Claim 5 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, wherein at least 90% of filamentous phages in the phage population have a length within 8 nm of the length value.

### Claim 6 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, wherein the length value is below 50 nm.

### Claim 7 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, wherein the length value is greater than 50 nm but less than 820 nm.

### Claim 8 (depends on 1)

The phage population of claim 1, wherein the phage population has a count of at least 1e14 pfu.

### Claim 9 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, further comprising an active agent attached to filamentous phages of the population.

### Claim 10 (depends on 9)

The homogeneous engineered filamentous phage population of claim 9, wherein the active agent is selected from the group consisting of a therapeutic agent, an imaging agent and a combination thereof.

### Claim 11 (depends on 10)

The homogeneous engineered filamentous phage population of claim 10, wherein the active agent is an imaging agent.

### Claim 12 (depends on 11)

The homogeneous engineered filamentous phage population of claim 11, wherein the imaging agent is a carbon nanostructure.

### Claim 13 (depends on 9)

A therapeutic and/or imaging method comprising administering to a subject the homogeneous engineered filamentous phage population of claim 9.

### Claim 14 (depends on 13)

The method of claim 13 for treating and/or imaging a tumor.

### Claim 15 (depends on 14)

The method of claim 14, wherein the tumor is a central nervous system tumor.

### Claim 16 (depends on 14)

The method of claim 14, wherein filamentous phages of the phage population comprise a tumor targeting moiety.

### Claim 17 (depends on 16)

The method of claim 16, wherein the tumor targeting moiety comprises chlorotoxin.

### Claim 18 (depends on 17)

The method of claim 17, wherein the chlorotoxin is a fluorescently labeled chlorotoxin.

### Claim 19 (depends on 17)

The method of claim 17, wherein the tumor is selected from glioma, medulloblastoma, neuroblastoma, melanoma, primitive neuroectodermal tumor, and small cell lung carcinoma.

### Claim 20 (depends on 13)

The method of claim 13, wherein the administering comprises intravascular administering.

### Claim 21 (depends on 13)

The method of claim 13, wherein the administering comprises implanting the homogeneous engineered filamentous phage population.

### Claim 22 (depends on 21)

The method of claim 21, wherein the said implanting is intracranial implanting.

### Claim 23 (depends on 1)

A pharmaceutical composition comprising the homogeneous engineered filamentous phage population of claim 1.

### Claim 24 (depends on 1)

An implant comprising the homogeneous engineered filamentous phage population of claim 1.

### Claim 25 (depends on 24)

The implant of claim 24, which is one or more of a scaffold, a hydrogel or a nanostructure.

### Claim 26

A phage comprising chlorotoxin expressed by a coat protein of the phage, wherein the phage is an engineered phage, which is a part of a homogeneous engineered phage population, wherein at least 70% of phages in the phage population have a length within 15% or within 8 nm of a length value, which is selected from 10 nm to less than the corresponding wild type phage length.

### Claim 27 (depends on 26)

The phage of claim 26, wherein the coat protein is pIII protein.

### Claim 28 (depends on 26)

The phage of claim 26, wherein at least 70% of phages in the phage population have a length within 15% of the length value.

### Claim 29 (depends on 26)

The phage of claim 26, wherein at least 70% of phages in the phage population have a length within 8 nm of the length value.

### Claim 30 (depends on 26)

The phage of claim 26, further comprising an imaging agent.

### Claim 31 (depends on 30)

The phage of claim 30, wherein the imaging agent comprises a carbon nanostructure.

### Claim 32 (depends on 26)

The phage of claim 26, wherein the chlorotoxin is a labeled chlorotoxin.

### Claim 33 (depends on 32)

The phage of claim 32, wherein the chlorotoxin is a fluorescently labeled chlorotoxin.

### Claim 34 (depends on 26)

A pharmaceutical composition comprising the phage of claim 26.

### Claim 35 (depends on 26)

An implant comprising the phage of claim 26.

### Claim 36 (depends on 35)

The implant of claim 35, which is one or more of a scaffold, a hydrogel or a nanostructure.

### Claim 37 (depends on 26)

A therapeutic and/or imaging method comprising administering to a subject the phage of claim 26.

### Claim 38 (depends on 37)

The method of claim 37 for treating and/or imaging a tumor.

### Claim 39 (depends on 38)

The method of claim 38, wherein the tumor is selected from glioma, medulloblastoma, neuroblastoma, melanoma, primitive neuroectodermal tumor, and small cell lung carcinoma.

### Claim 40 (depends on 37)

The method of claim 37, wherein said administering is performed intravascularly.

### Claim 41 (depends on 40)

The method of claim 40, wherein the phage penetrates a blood-brain barrier in the subject upon said administering.

### Claim 42 (depends on 37)

The method of claim 37, wherein said administering is performed intracranially.

### Claim 43 (depends on 37)

The method of claim 37, wherein said administering comprises implanting the phage.

### Claim 44 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, which is a homogeneous engineered Ff phage population.

### Claim 45 (depends on 44)

The homogeneous engineered filamentous phage population of claim 44, which is a homogeneous engineered M13 phage population.

### Claim 46 (depends on 26)

The phage of claim 26, wherein the engineered phage is an engineered filamentous phage.

### Claim 47 (depends on 46)

The phage of claim 46, wherein the engineered filamentous phage is an engineered Ff phage.

### Claim 48 (depends on 46)

The phage of claim 46, wherein the engineered filamentous phage is an engineered M13 phage.

### Claim 49 (depends on 45)

The homogeneous engineered filamentous phage population of claim 45, wherein the selected length value is from 10 nm to 850 nm.

### Claim 50 (depends on 48)

The phage of claim 48, wherein the engineered filamentous phage is an engineered M13 phage, wherein the selected length value is from 10 nm to 850 nm.

### Claim 51 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, wherein the selected length value is from 10 nm to 150 nm.

### Claim 52 (depends on 46)

The phage of claim 46, wherein the selected length value is from 10 nm to 150 nm.

### Claim 53 (depends on 1)

The homogeneous engineered filamentous phage population of claim 1, wherein the selected length value is from 50 nm to 300 nm.

### Claim 54 (depends on 46)

The phage of claim 46, wherein the selected length value is from 50 nm to 300 nm.