CA-3286074-A1 - RECOMBINANT HERPES SIMPLEX VIRUS WITH IL-12 EXPRESSION

Abstract

An object of the present invention is to develop a novel armed oncolytic herpes simplex virus (HSV) using G47Δ as the backbone. There is provided a pharmaceutical composition comprising a recombinant herpes simplex virus (HSV) with interleukin 12 (IL-12) expression, the recombinant HSV having a gene encoding a fusion polypeptide in which the 35 kDa light chain (p35) and the 40 kDa heavy chain (p40) of IL-12 are linked via two or more elastin motifs, and having the following characteristics (a) to (c): (a) the ICP6 gene is deleted or inactivated; (b) the γ34.5 gene is deleted or inactivated; and (c) the α47 gene is deleted or inactivated.

Inventors

• Tomoki Todo
• Hiroshi Fukuhara

Assignees

• Tomoki Todo

Dates

Publication Date

20260301

Application Date

20240318

Priority Date

20230317

Claims (1)

1. <pat:ClaimStatement>Claims</pat:ClaimStatement> <pat:Claims com:id="claims"> <pat:Claim com:id="CLM-00001"> <pat:ClaimNumber>1</pat:ClaimNumber> <pat:ClaimText>1. A pharmaceutical composition comprising a recombinant herpes simplex virus (HSV) with interleukin 12 (IL-12) expression, the recombinant HSV comprising a gene encoding a fusion polypeptide in which the 35 kDa light chain (p35) and the 40 kDa heavy chain (p40) of IL-12 are linked via two or more elastin motifs, and having the following characteristics (a) to (c): (a) the ICP6 gene is deleted or inactivated, (b) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>γ</mi><annotation encoding="application/x-tex">\gamma</annotation></semantics></math>34.5 gene is deleted or inactivated, and (c) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>α</mi><mn>47</mn></mrow><annotation encoding="application/x-tex">\alpha 47</annotation></semantics></math> gene is deleted or inactivated. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00002"> <pat:ClaimNumber>2</pat:ClaimNumber> <pat:ClaimText>2. The pharmaceutical composition according to claim 1, wherein the fusion polypeptide comprises p40, two or more elastin motifs, and p35 linked in this order from the N-terminal side. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00003"> <pat:ClaimNumber>3</pat:ClaimNumber> <pat:ClaimText>3. The pharmaceutical composition according to claim 1, wherein the gene encoding the fusion polypeptide is inserted into the ICP6 locus. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00004"> <pat:ClaimNumber>4</pat:ClaimNumber> <pat:ClaimText>4. The pharmaceutical composition according to claim 1, wherein the gene encoding the fusion peptide is any of the following polynucleotides: (i) a polynucleotide consisting of the sequence of SEQ ID NO: 1; (ii) a polynucleotide consisting of a sequence having a sequence identity of 90% or higher to the sequence of SEQ ID NO: 1 and encoding a polypeptide capable of forming an active IL-12; (iii) a polynucleotide encoding a polypeptide consisting of the sequence of SEQ ID NO: 2; (iv) a polynucleotide encoding a polypeptide consisting of a sequence having a sequence identity of 90% or higher to the sequence of SEQ ID NO: 2 and capable of forming an active IL-12; (v) a polynucleotide consisting of the sequence of SEQ ID NO: 3; (vi) a polynucleotide consisting of a sequence having a sequence identity of 90% or higher to the sequence of SEQ ID NO: 3 and encoding a polypeptide capable of forming an active IL-12; (vii) a polynucleotide encoding a polypeptide consisting of the sequence of SEQ ID NO: 4; and (viii) a polynucleotide encoding a polypeptide consisting of a sequence having a sequence identity of 90% or higher to the sequence of SEQ ID NO: 4 and capable of forming an active IL-12. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00005"> <pat:ClaimNumber>5</pat:ClaimNumber> <pat:ClaimText>5. The pharmaceutical composition according to claim 1, wherein the recombinant HSV is derived from <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>G</mi><mn>47</mn><mi mathvariant="normal">Δ</mi></mrow><annotation encoding="application/x-tex">G47\Delta</annotation></semantics></math>. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00006"> <pat:ClaimNumber>6</pat:ClaimNumber> <pat:ClaimText>6. The pharmaceutical composition according to any one of claims 1 to 5, which is used for enhancing the effect of IL-12 in a treatment of a tumor. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00007"> <pat:ClaimNumber>7</pat:ClaimNumber> <pat:ClaimText>7. The pharmaceutical composition according to claim 5, which is for local injection. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00008"> <pat:ClaimNumber>8</pat:ClaimNumber> <pat:ClaimText>8. A method for enhancing the effect of IL-12 in a treatment of a tumor, the method comprising administering the pharmaceutical composition comprising a recombinant HSV with IL-12 expression according to any one of claims 1 to 5 to a subject in need of the enhancement. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00009"> <pat:ClaimNumber>9</pat:ClaimNumber> <pat:ClaimText>9. A pharmaceutical composition comprising a recombinant HSV with IL-12 expression, the recombinant HSV comprising a gene encoding IL-12, and having the following characteristics (a) to (c), which induces IFNy production through IL-12 expression without decreasing the replication ability of the recombinant HSV: (a) the ICP6 gene is deleted or inactivated, (b) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>γ</mi><annotation encoding="application/x-tex">\gamma</annotation></semantics></math>34.5 gene is deleted or inactivated, and (c) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>α</mi><annotation encoding="application/x-tex">\alpha</annotation></semantics></math>47 gene is deleted or inactivated. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00010"> <pat:ClaimNumber>10</pat:ClaimNumber> <pat:ClaimText>10. A pharmaceutical composition for intravenous administration, comprising a recombinant HSV with IL-12 expression, the recombinant HSV comprising a gene encoding IL-12, and having the following characteristics (a) to (c): (a) the ICP6 gene is deleted or inactivated, (b) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>γ</mi><annotation encoding="application/x-tex">\gamma</annotation></semantics></math>34.5 gene is deleted or inactivated, and (c) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>α</mi><annotation encoding="application/x-tex">\alpha</annotation></semantics></math>47 gene is deleted or inactivated. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00011"> <pat:ClaimNumber>11</pat:ClaimNumber> <pat:ClaimText>11. A pharmaceutical composition for intratumoral administration, comprising a recombinant HSV with IL-12 expression, the recombinant HSV comprising a gene encoding IL-12, and the following characteristics (a) to (c), which elicits systemic anti- tumor immunity, and thereby attains either the suppression of metastasis or the suppression of growth of a tumor not receiving direct intratumoral administration, or both: (a) the ICP6 gene is deleted or inactivated, (b) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>γ</mi><annotation encoding="application/x-tex">\gamma</annotation></semantics></math>34.5 gene is deleted or inactivated, and (c) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>α</mi><mn>47</mn></mrow><annotation encoding="application/x-tex">\alpha 47</annotation></semantics></math> gene is deleted or inactivated. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00012"> <pat:ClaimNumber>12</pat:ClaimNumber> <pat:ClaimText>12. A pharmaceutical composition for treating a tumor requiring increased infiltration of immune cells comprising a recombinant HSV with IL-12 expression, the recombinant HSV comprising a gene encoding IL-12, and having the following characteristics (a) to (c): (a) the ICP6 gene is deleted or inactivated, (b) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>γ</mi><annotation encoding="application/x-tex">\gamma</annotation></semantics></math>34.5 gene is deleted or inactivated, and (c) the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>α</mi><mn>47</mn></mrow><annotation encoding="application/x-tex">\alpha 47</annotation></semantics></math> gene is deleted or inactivated. </pat:ClaimText> </pat:Claim> <pat:Claim com:id="CLM-00013"> <pat:ClaimNumber>13</pat:ClaimNumber> <pat:ClaimText>13. The pharmaceutical composition according to any one of claims 9 to 12, wherein the recombinant HSV is the recombinant HSV defined in any one of claims 1 to 5. </pat:ClaimText> </pat:Claim> </pat:Claims>

Description

Specification Title of the Invention: Recombinant herpes simplex virus with IL-12 expression Technical Field [0001] The present invention relates to an oncolytic herpes virus. More specifically, the present invention relates to a function-added oncolytic herpes virus that enhances therapeutic effect of viral therapy. Background Art [0002] Oncolytic herpes simplex virus type 1 (HSV-1) is an effective tool for the treatment of cancer (Non-patent documents 1 and 2). The first oncolytic HSV-1 having a deletion in the thymidine kinase gene was reported in 1991 (Non-patent document 3). Since then, various oncolytic HSV-1s have been validated in clinical trials. T-VEC (Talimogene laherparepvec), an oncolytic HSV-1 with mutations in the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>γ</mi><annotation encoding="application/x-tex">\gamma</annotation></semantics></math>34.5 and <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>α</mi><annotation encoding="application/x-tex">\alpha</annotation></semantics></math>47 genes, was approved as a drug for inoperable melanoma in the United States and Europe in 2015 (Non-patent document 4). G207, one of the oncolytic HSV-1 viruses first used in clinical trials, has deletions in both copies of the <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>γ</mi><annotation encoding="application/x-tex">\gamma</annotation></semantics></math>34.5 gene and a lacZ insertion that inactivates the ICP6 gene (Non-patent document 5). <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>G</mi><mn>47</mn><mi mathvariant="normal">Δ</mi></mrow><annotation encoding="application/x-tex">G47\Delta</annotation></semantics></math> was constructed by further introducing a deletion into the US11 promoter overlapping with the α47 gene of the G207 genome (Non-patent document 6, and Patent document 1). The efficacies of <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>G</mi><mn>47</mn><mi mathvariant="normal">Δ</mi></mrow><annotation encoding="application/x-tex">G47\Delta</annotation></semantics></math> against a variety of cancers have been demonstrated (Non-patent documents 7 to 16). Safety of <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>G</mi><mn>47</mn><mi mathvariant="normal">Δ</mi></mrow><annotation encoding="application/x-tex">G47\Delta</annotation></semantics></math> has been verified in multiple clinical trials conducted in Japan, including those for glioblastoma (UMIN00002661, Non-patent document 17), castration-resistant prostate cancer (UMIN 000010463), metastatic prostate cancer (jRCTs033210603), olfactory neuroblastoma (UMIN000011636, jRCTs03180325), and malignant pleural mesothelioma (UMIN000034063, jRCTs03180326). Recently, a phase II clinical trial for glioblastoma (UMIN000015995) led to the approval of <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>G</mi><mn>47</mn><mi mathvariant="normal">Δ</mi></mrow><annotation encoding="application/x-tex">G47\Delta</annotation></semantics></math> as a new drug for malignant glioma in Japan (Non-patent document 18). [0003] As the next generation of oncolytic HSV-1, function-added G47<math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi mathvariant="normal">Δ</mi><annotation encoding="application/x-tex">\Delta</annotation></semantics></math> incorporating various exogenous genes into the basic backbone of <math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mrow><mi>G</mi><mn>47</mn><mi mathvariant="normal">Δ</mi></mrow><annotation encoding="application/x-tex">G47\Delta</annotation></semantics></math>, for example, a vascular endothelial growth factor (VEGF)-expressing and swelling-suppressive oncolytic virus incorporating a gene encoding a VEGF antagonist (Patent document 2), is under development. [0004] By the way, interleukin-12 (IL-12) is a promising tool for cancer therapy, and several types of mouse IL-12 have been confirmed to be expressed by oncolytic HSV-1 (Non-patent documents 19 to 22). In general, tumor formation and growth are highly dependent on the host's inability to elicit strong anti-tumor immune responses and formation of new blood vessels to feed the tumor. IL-12 can target these two processes. IL-12 is a cytokine that activates a variety of immune cells, including NK cells and T cells. In fact, IFNy is induced and secreted by these cells. [0005] Functional IL-12, p70, is a heterodimeric glycoprotein composed of two subunits, a 35 kDa light chain (p35) and a 40 kDa heavy chain (p40) (Non-patent documents 23 and 24). The p70-IL-12Rβ1 complex binds to another second p70, and the resulting complex preforms a binding site with high affinity for IL-12R<math xmlns="http://www.w3.org/1998/Math/MathML"><semantics><mi>β</mi><annotation encoding="application/x-tex">\beta</annotation></semantics></math>2. The interaction between p40 and IL-12Rβ1 is required to stabilize the p70-IL-12Rβ1 complex (Non-paten