# US 2023/0272402 A1 · Small rna medicament for prevention and treatment of inflammation-related diseases and combination thereof

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- Number: US 2023/0272402 A1
- Document ID: US20230272402A1
- Status: Published application
- Kind: A1
- Application number: 17/417,321
- Filed: 2018-12-25
- Published: 2023-08-31
- Earliest claimed: 2018-12-25
- Assignees: Beijing Baishihekang Pharmaceutical Technology (Bsjpharma) Co., Ltd
- Applicants: Institute of Basic Medical Sciences Chinese Academy of Medical Sciences
- Inventors: Chengyu JIANG; Dandan ZHAO; Yuhao QIN; Cong ZHANG; Yexuan LIN
- Web page: https://patentgrep.com/patent/US20230272402A1

## CPC

- C12N 15/1136: {against growth factors, growth regulators, cytokines, lymphokines or hormones}
- A61P 29/00: Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
- A61P 35/00: Antineoplastic agents
- C12N 15/85: for animal cells
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2330/10: naturally occurring
- C12N 2330/30: chemically synthesised
- C12N 2330/50: Biochemical production, i.e. in a transformed host cell
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Provided is a small RNA, a composition comprising the small RNA, a method for using same and use of same. The small RNA or composition can inhibit the ability of any one or more of pathways or genes listed in Table 3, or decrease or down regulate the expression level of IL-1 beta, IL-6, and/or TNF-alpha in vitro or in vivo, or treat or prevent IL-1 beta, IL-6, and/or TNF-alpha related diseases and/or increase cell viability in a subject.

## Claims

### Claim 1

A small RNA comprising:
(A) The sequence shown in any one of SEQ ID NO. 1-222, or complementary sequence thereof;
(B) A sequence with at least 80%-98% identity with the sequence shown in (A);
(C) A sequence that hybridizes to the sequence shown in (A);
(D) A sequence obtained from the sequence shown in (A) by addition, deletion, replacement or insertion of one or more; or
(E) A precursor or modified variant of the sequence shown in (A), (B), (C) or (D).

### Claim 2 (depends on 1)

The small RNA according to claim 1, which has an ability to prevent and/or treat IL-1beta, IL-6 or/and TNF-alpha related diseases, and/or an ability to improve cell survival rate.

### Claim 3 (depends on 2)

The small RNA according to claim 2, wherein the small RNA has an ability to reduce or down-regulate the expression level of IL-1beta, IL-6 or/and TNF-alpha and/or has an ability to rescue cell death caused by a virus infection,
wherein the small RNA has an ability to reduce or down-regulate the expression level of an inflammatory factor selected form any one of IL-1beta, IL-6 and TNF-alpha,
wherein the IL-1beta, IL-6 or/and TNF-alpha related disease is selected from the group consisting of: pneumonia, myocarditis, acute and chronic gastritis, acute and chronic enteritis, acute and chronic hepatitis, acute and chronic nephritis, dermatitis, encephalitis, lymphitis, conjunctivitis, keratitis, iridocyclitis, tympanitis, allergic rhinitis, asthma, pulmonary fibrosis, chronic obstructive pulmonary disease, allergic dermatitis, sickle cell disease, multiple sclerosis, systemic lupus erythematosus, lupus nephritis, lung cancer, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, cervical cancer, breast cancer, leukemia, multiple myeloma, diabetes and gout.

### Claim 4 (depends on 1, 2, 3)

The small RNA according to any one of claims 1 to 3, wherein the small RNA is in a double-stranded or single-stranded form or a double-stranded and single-stranded hybrid form.

### Claim 5 (depends on 1, 2, 3)

The small RNA according to any one of claims 1 to 3 wherein the small RNA is a non-natural small RNA.

### Claim 6 (depends on 1, 2, 3)

The small RNA according to any one of claims 1 to 3 wherein the non-natural small RNA is a small RNA obtained through artificial synthesis or expression of an artificial vector.

### Claim 7 (depends on 1, 2, 3)

A nucleic acid sequence or a construct comprising a sequence encoding the small RNA according to any one of claims 1 to 3, wherein the construct is a viral construct.

### Claim 8 (depends on 7)

A recombinant virus comprising the nucleic acid sequence or construct according to claim 7, wherein the recombinant virus is a retrovirus.

### Claim 9 (depends on 1, 2, 3)

An expression vector comprising a sequence encoding the small RNA according to any one of claims 1 to 3.

### Claim 10 (depends on 7)

A cell comprising the nucleic acid sequence or construct according to claim 7.

### Claim 11 (depends on 1, 2, 3)

A method for expressing the small RNA according to any one of claims 1 to 3, comprising:
expressing a cell comprising a nucleic acid sequence or a construct, the nucleic acid sequence comprising a sequence encoding the small RNA; and
recovering the small RNA.

### Claim 12 (depends on 1, 2, 3)

A pharmaceutical composition comprising at least one of:
one or more small RNA(s) according to any one of claims 1 to 3,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s), and
a cell comprising the nucleic acid sequence or construct,
wherein said pharmaceutical composition is prepared for oral administration, intravenous administration, subcutaneous administration, intramuscular administration, intraarterial administration, intraperitoneal administration, intrapulmonary administration, intracerebrospinal administration, intraarticular administration, intrasynovial administration, intrathecal administration, intralesional administration, or administration by inhalation.

### Claim 13 (depends on 12)

The pharmaceutical composition according to claim 12, comprising any one or more of mixture(s) selected from the group consisting of:
Mixture-1: BZL-sRNA-20, BZL-sRNA-2, BZL-sRNA-6, BZL-sRNA-9, BZL-sRNA-40, BZL-sRNA-41:
Mixture-2: BZL-sRNA-20, BZL-sRNA-4, BZL-sRNA-8, BZL-sRNA-12, BZL-sRNA-21, BZL-sRNA-31;
Mixture-3: BZL-sRNA-20, BZL-sRNA-33, BZL-sRNA-34, BZL-sRNA-35, BZL-sRNA-38, BZL-sRNA-39;
Mixture-4: BZL-sRNA-20, BZL-sRNA-11, BZL-sRNA-13, BZL-sRNA-19, BZL-sRNA-23, BZL-sRNA-28:
Mixture-5: BZL-sRNA-20, BZL-sRNA-29, BZL-sRNA-36, BZL-sRNA-37, BZL-sRNA-42:
Mixture-6: BZL-sRNA-20, CHu-sRNA-23, CHu-sRNA-27, CHu-sRNA-31, CHu-sRNA-35, CHu-sRNA-41;
Mixture-7: BZL-sRNA-20, CHu-sRNA-46, CHu-sRNA-47, CHu-sRNA-48, CHu-sRNA-49, CHu-sRNA-50;
Mixture-8: BZL-sRNA-20, CHu-sRNA-52, CHu-sRNA-53, CHu-sRNA-54, CHu-sRNA-6, CHu-sRNA-43:
Mixture-9: BZL-sRNA-20, CHu-sRNA-51, CHu-sRNA-55, CHu-sRNA-5, CHu-sRNA-8, CHu-sRNA-10:
Mixture-10: BZL-sRNA-20, CHu-sRNA-21, CHu-sRNA-24, CHu-sRNA-28, CHu-sRNA-29, CHu-sRNA-30, CHu-sRNA-42;
Mixture-11: BZL-sRNA-20, LQi-sRNA-7, DDi-sRNA-1, HQi-sRNA-1, XKC-sRNA-4, XKC-sRNA-8;
Mixture-12: BZL-sRNA-20, IYH-sRNA-1, LOi-sRNA-5, LOi-sRNA-8, XKC-sRNA-2, XKC-sRNA-7:
Mixture-13: BZL-sRNA-20, YXC-sRNA-15, YXC-sRNA-30, YXC-sRNA-32, YXC-sRNA-33, YXC-sRNA-34:
Mixture-14: BZL-sRNA-20, YXC-sRNA-35, YXC-sRNA-38, YXC-sRNA-39, YXC-sRNA-40, YXC-sRNA-43;
Mixture-15: BZL-sRNA-20, YXC-sRNA-46, YXC-sRNA-47, YXC-sRNA-50, YXC-sRNA-51, YXC-sRNA-53;
Mixture-16: BZL-sRNA-20, YXC-sRNA-56, YXC-sRNA-62, YXC-sRNA-64, YXC-sRNA-70, YXC-sRNA-71:
Mixture-17: BZL-sRNA-20, YXC-sRNA-72, YXC-sRNA-75, YXC-sRNA-79, YXC-sRNA-82:
Mixture-18: BZL-sRNA-20, YXC-sRNA-1, YXC-sRNA-2, YXC-sRNA-3, YXC-sRNA-9, YXC-sRNA-10;
Mixture-19: BZL-sRNA-20, YXC-sRNA-12, YXC-sRNA-13, YXC-sRNA-16, YXC-sRNA-19, YXC-sRNA-24;
Mixture-20: BZL-sRNA-20, YXC-sRNA-25, YXC-sRNA-26, YXC-sRNA-28, YXC-sRNA-29, YXC-sRNA-3 I:
Mixture-21: BZL-sRNA-20, YXC-sRNA-36, YXC-sRNA-37, YXC-sRNA-41, YXC-sRNA-42, YXC-sRNA-44;
Mixture-22: BZL-sRNA-20, YXC-sRNA-48, YXC-sRNA-49, YXC-sRNA-54, YXC-sRNA-55, YXC-sRNA-57;
Mixture-23: BZL-sRNA-20, YXC-sRNA-58, YXC-sRNA-60, YXC-sRNA-66, YXC-sRNA-68, YXC-sRNA-69;
Mixture-24: BZL-sRNA-20, YXC-sRNA-73, YXC-sRNA-80, YXC-sRNA-83;
Mixture-25: BZL-sRNA-20, PGY-sRNA-21, PGY-sRNA-22, PGY-sRNA-23, PGY-sRNA-25, PGY-sRNA-26;
Mixture-26: BZL-sRNA-20, PGY-sRNA-28, PGY-sRNA-29, PGY-sRNA-30, PGY-sRNA-32;
Mixture-27: BZL-sRNA-20, CXL-sRNA-30, PGY-sRNA-27, PGY-sRNA-31, PGY-sRNA-24;
Mixture-28: BZL-sRNA-20, BZL-sRNA-1, BZL-sRNA-3, BZL-sRNA-5, BZL-sRNA-7, BZL-sRNA-10;
Mixture-29: BZL-sRNA-20, BZL-sRNA-14, BZL-sRNA-15, BZL-sRNA-16, BZL-sRNA-17, BZL-sRNA-18, BZL-sRNA-30;
Mixture-30: BZL-sRNA-20, BZL-sRNA-22, BZL-sRNA-24, BZL-sRNA-25, BZL-sRNA-26, BZL-sRNA-27, BZL-sRNA-32;
Mixture-31: BZL-sRNA-20, CHu-sRNA-1, CHu-sRNA-2, CHu-sRNA-3, CHu-sRNA-4, CHu-sRNA-7;
Mixture-32: BZL-sRNA-20, CHu-sRNA-9, CHu-sRNA-11, CHu-sRNA-12, CHu-sRNA-13, CHu-sRNA-14;
Mixture-33: BZL-sRNA-20, CHu-sRNA-15, CHu-sRNA-16, CHu-sRNA-17, CHu-sRNA-18, CHu-sRNA-19;
Mixture-34: BZL-sRNA-20, CHu-sRNA-20, CHu-sRNA-22, CHu-sRNA-25, CHu-sRNA-26, CHu-sRNA-32;
Mixture-35: BZL-sRNA-20, CHu-sRNA-33, CHu-sRNA-34, CHu-sRNA-36, CHu-sRNA-37, CHu-sRNA-38;
Mixture-36: BZL-sRNA-20, CHu-sRNA-39, CHu-sRNA-40, CHu-sRNA-44, CHu-sRNA-45;
Mixture-37: BZL-sRNA-20, HQi-sRNA-2, LQi-sRNA-1, LQi-sRNA-2, LQi-sRNA-3, LQi-sRNA-4;
Mixture-38: BZL-sRNA-20, LQi-sRNA-6, XKC-sRNA-1, XKC-sRNA-3, XKC-sRNA-5, XKC-sRNA-6;
Mixture-39: BZL-sRNA-20, YXC-sRNA-4, YXC-sRNA-5, YXC-sRNA-6, YXC-sRNA-7, YXC-sRNA-8;
Mixture-40: BZL-sRNA-20, YXC-sRNA-11, YXC-sRNA-14, YXC-sRNA-17, YXC-sRNA-18, YXC-sRNA-20;
Mixture-41: BZL-sRNA-20, YXC-sRNA-2 I, YXC-sRNA-22, YXC-sRNA-23, YXC-sRNA-27, YXC-sRNA-45;
Mixture-42: BZL-sRNA-20 YXC-sRNA-52 YXC-sRNA-59 YXC-sRNA-61,YXC-sRNA-63, YXC-sRNA-65; and
Mixture-43: BZL-sRNA-20, YXC-sRNA-67, YXC-sRNA-74, YXC-sRNA-81, YXC-sRNA-76, YXC-sRNA-77,YXC-sRNA-78,

### Claim 14 (depends on 13)

The pharmaceutical composition according to claim 13, wherein in the pharmaceutical composition, the molar concentration ratio of the small RNA comprising the sequence shown in SEQ ID NO. 20 to other small RNA(s) in the composition is about 2:1.

### Claim 15

(canceled)

### Claim 16 (depends on 1, 2, 3)

A kit comprising:
at least one of:
one or more small RNA(s) according to any one of claims 1 to 3,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s), and
a cell comprising the nucleic acid sequence or construct.

### Claim 17 (depends on 1, 2, 3)

A method for inhibiting inflammatory factors and/or virus infection in vitro or in vivo, comprising administering to a cell or a subject at least one of:
one or more small RNA(s) according to any one of claims 1 to 3,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s),
a cell comprising the nucleic acid sequence or construct; and
a pharmaceutical composition comprising the one or more small RNA(s), the recombinant virus, the expression vector, or the cell.

### Claim 18 (depends on 1, 2, 3)

A method for reducing or down-regulating the expression level of IL-1beta, IL-6 or/and TNF-alpha in vitro or in vivo and/or improving cell survival rate, which comprises administering to a cell or a subject at least one of:
one or more small RNA(s) according to any one of claims 1 to 3,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s),
a cell comprising the nucleic acid sequence or construct; and
a pharmaceutical composition comprising the one or more small RNA(s), the recombinant virus, the expression vector, or the cell.

### Claim 19 (depends on 18)

The method according to claim 18, wherein the cell survival rate is the cell survival rate in virus infection, wherein said cell survival rate is improved by rescuing the cell death caused by the virus infection.

### Claim 20 (depends on 1, 2, 3)

A method for treating or preventing IL-1beta, IL-6 or/and TNF-alpha related diseases and/or virus infection in a subject, which comprises administering to a subject at least one of:
one or more small RNA(s) according to any one of claims 1 to 3,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s), a cell comprising the nucleic acid sequence or construct; and
a pharmaceutical composition comprising the one or more small RNA(s), the recombinant virus, the expression vector, or the cell,
wherein the IL-1beta, IL-6 or/and TNF-alpha related disease is selected from the group consisting of: pneumonia, myocarditis, acute and chronic gastritis, acute and chronic enteritis, acute and chronic hepatitis, acute and chronic nephritis, dermatitis, encephalitis, lymphitis, conjunctivitis, keratitis, iridocyclitis, tympanitis, allergic rhinitis, asthma, pulmonary fibrosis, chronic obstructive pulmonary disease, allergic dermatitis, sickle cell disease, multiple sclerosis, systemic lupus erythematosus, lupus nephritis, lung cancer, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, cervical cancer, breast cancer, leukemia, multiple myeloma, diabetes, and gout.

### Claim 21 (depends on 1, 2, 3)

A method for reducing or down-regulating the expression level of IL-1beta, IL-6 or/and TNF-alpha in vitro or in vivo and/or improving cell survival rate and/or treating or preventing IL-1beta, IL-6 or TNF-alpha related diseases and/or virus infection in a subject, comprising administering to the subject at least one of:
one or more small RNA(s) according to any one of claims 1 to 3,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s),
a cell comprising the nucleic acid sequence or construct; and
a pharmaceutical composition comprising the one or more small RNA(s), the recombinant virus, the expression vector, or the cell,
wherein the IL-1beta, IL-6 or/and TNF-alpha related disease is selected from the group consisting of: pneumonia, myocarditis, acute and chronic gastritis, acute and chronic enteritis, acute and chronic hepatitis, acute and chronic nephritis, dermatitis, encephalitis, lymphitis, conjunctivitis, keratitis, iridocyclitis, tympanitis, allergic rhinitis, asthma, pulmonary fibrosis, chronic obstructive pulmonary disease, allergic dermatitis, sickle cell disease, multiple sclerosis, systemic lupus erythematosus, lupus nephritis, lung cancer, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, cervical cancer, breast cancer, leukemia, multiple myeloma, diabetes, and gout.

### Claim 22 (depends on 1, 2, 3)

A method for preparing a medicament for reducing or down-regulating the expression level of IL-1beta, IL-6 or/and TNF-alpha in vitro or in vivo and/or improving cell survival rate and/or treating or preventing IL-1beta, IL-6 or/and TNF-alpha related diseases and/or virus infection in a subject, comprising administering to the subject at least one of:
one or more small RNA(s) according to any one of claims 1 to 3,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s),
a cell comprising the nucleic acid sequence or construct; and
a pharmaceutical composition comprising the one or more small RNA(s), the recombinant virus, the expression vector, or the cell,
wherein the IL-1beta, IL-6 or/and TNF-alpha related disease is selected from the group consisting of: pneumonia, myocarditis, acute and chronic gastritis, acute and chronic enteritis, acute and chronic hepatitis, acute and chronic nephritis, dermatitis, encephalitis, lymphitis, conjunctivitis, keratitis, iridocyclitis, tympanitis, allergic rhinitis, asthma, pulmonary fibrosis, chronic obstructive pulmonary disease, allergic dermatitis, sickle cell disease, multiple sclerosis, systemic lupus erythematosus, lupus nephritis, lung cancer, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, cervical cancer, breast cancer, leukemia, multiple myeloma, diabetes, and gout.

### Claim 23 (depends on 1, 2, 3)

A reagent for detecting at least one of following:
the small RNA according to any one of claims 1 to 3,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s),
a cell comprising the nucleic acid sequence or construct; and
a pharmaceutical composition comprising the one or more small RNA(s), the recombinant virus, the expression vector, or the cell,
wherein the reagent is a primer and/or a probe.

### Claim 24 (depends on 23)

A kit comprising the reagent according to claim 23.

### Claim 25 (depends on 1, 2, 3)

A method for detecting whether cells from different sources contain the small RNA according to any one of claims 1 to 3, comprising providing:
(i) a reagent for detecting at least one of following:
the small RNA,
a nucleic acid sequence or a construct comprising the one or more small RNA(s),
a recombinant virus comprising the nucleic acid sequence or construct,
an expression vector comprising a sequence encoding the one or more small RNA(s),
a cell comprising the nucleic acid sequence or construct; and
a pharmaceutical composition comprising the one or more small RNA(s), the recombinant virus, the expression vector, or the cell,
wherein the reagent is a primer and/or a probe; or
(ii) a kit comprising the reagent;
wherein the cell is a plant cell.