Treatment of diabetic retinopathy with whole human post-translationally modified anti-VEGF Fab

Abstract

Described are methods for using a fully human post-translationally modified (HuPTM) monoclonal antibody ("mAb") or antigen binding fragment of mAb against human vascular endothelial growth factor ("hVEGF"), such as, for example, an antigen binding fragment of mAb. The present invention relates to compositions and methods for delivery of a fully human glycosylated (HuGly) anti-hVEGF antigen binding fragment to the retinal / vitreous humor in the eye of a human subject diagnosed with diabetic retinopathy. The invention relates to compositions and methods for the delivery of a fully human glycosylated (HuGly) anti-hVEGF antigen binding fragment to the retinal / vitreous humor in the eye of a human subject diagnosed with diabetic retinopathy.

Description

[0001] Cross References to Related Applications

[0002] This application claims U.S. Provisional Application No. 62 / 891,799, filed August 26, 2019, U.S. Provisional Application No. 62 / 902,352, filed September 18, 2019, and U.S. Provisional Application No. 63, filed April 2, 2020 / 004,258, the contents of which are incorporated herein by reference in their entirety.

[0003] References to Sequence Listings Submitted Electronically

[0004] This application incorporates by reference the Sequence Listing entitled "12656-127-228_Sequence_Listing.TXT," which was created on August 12, 2020 and is 97,447 bytes in size, as a text file filed with this application. technical field

[0005] A fully human post-translationally modified (HuPTM) monoclonal antibody ("mAb") or an antigen-binding fragment of a mAb directed against vascular endothelial growth factor ("VEGF") is described—for example, a fully human glycosylated (HuGly) Anti-VEGF Antigen-Binding Fragment - delivered to the ...

Examples

example 2

[0416] 6.2 Example 2: Ranibizumab cDNA-based vector

[0417] A ranibizumab Fab cDNA-based vector comprising a transgene comprising the ranibizumab Fab light and heavy chain cDNAs (not encoding the portion of SEQ ID NO. 12 and 13 of the signal peptide, respectively) was constructed. Transgenes also include nucleic acids comprising a signal peptide selected from the group listed in Table 1. The nucleotide sequences encoding the light and heavy chains are separated by an IRES element or 2A cleavage site to generate a bicistronic vector. Optionally, the vector additionally includes a hypoxia-inducible promoter.

example 3

[0418] 6.3 Example 3: Vectors based on hyperglycosylated bevacizumab Fab cDNA

[0419] A hyperglycosylated bevacizumab Fab cDNA-based vector comprising a transgene comprising the bevacizumab Fab of the light and heavy chain cDNA sequences (SEQ ID NO. 10 and 11, respectively) was constructed. A portion having a mutation in the sequence encoding one or more of the following mutations: L118N (heavy chain), E195N (light chain) or Q160N or Q160S (light chain). Transgenes also include nucleic acids comprising a signal peptide selected from the group listed in Table 1. The nucleotide sequences encoding the light and heavy chains are separated by an IRES element or 2A cleavage site to generate a bicistronic vector. Optionally, the vector additionally includes a hypoxia-inducible promoter.

[0420] 6.4 Example 4: Hyperglycosylated ranibizumab cDNA based vector

[0421] A vector based on the hyperglycosylated ranibizumab Fab cDNA comprising the transgene comprising the ranibizumab li...

example 7

[0476] 6.7 Example 7: Open-label Phase 2a Dose Evaluation of Construct II Gene Therapy in Participants with Diabetic Retinopathy

[0477] This example is an updated version of Example 6 and provides an overview of the Phase 2a dose evaluation of Construct II gene therapy in participants with diabetic retinopathy (DR). Continued stable expression of the construct II transgene product following a single gene therapy treatment for DR could potentially reduce the treatment burden of currently available therapies while preserving vision, its favorable benefit:risk spectrum. The current proof-of-concept study was designed to evaluate the safety and efficacy of Construct II gene therapy at 2 different dose levels in participants with DR.

[0478] 6.7.1 Purpose and Endpoints

[0479] Table 6: Primary Purposes and Endpoints and Secondary Purposes and Endpoints

[0480]

[0481]

[0482]

[0483] AAV8 = adeno-associated virus serotype 8; AE = adverse event; CI-DME = centrally...

[0001] Cross References to Related Applications

[0002] This application claims U.S. Provisional Application No. 62 / 891,799, filed August 26, 2019, U.S. Provisional Application No. 62 / 902,352, filed September 18, 2019, and U.S. Provisional Application No. 63, filed April 2, 2020 / 004,258, the contents of which are incorporated herein by reference in their entirety.

[0003] References to Sequence Listings Submitted Electronically

[0004] This application incorporates by reference the Sequence Listing entitled "12656-127-228_Sequence_Listing.TXT," which was created on August 12, 2020 and is 97,447 bytes in size, as a text file filed with this application. technical field

[0005] A fully human post-translationally modified (HuPTM) monoclonal antibody ("mAb") or an antigen-binding fragment of a mAb directed against vascular endothelial growth factor ("VEGF") is described—for example, a fully human glycosylated (HuGly) Anti-VEGF Antigen-Binding Fragment - delivered to the ...

Examples

example 2

[0416] 6.2 Example 2: Ranibizumab cDNA-based vector

[0417] A ranibizumab Fab cDNA-based vector comprising a transgene comprising the ranibizumab Fab light and heavy chain cDNAs (not encoding the portion of SEQ ID NO. 12 and 13 of the signal peptide, respectively) was constructed. Transgenes also include nucleic acids comprising a signal peptide selected from the group listed in Table 1. The nucleotide sequences encoding the light and heavy chains are separated by an IRES element or 2A cleavage site to generate a bicistronic vector. Optionally, the vector additionally includes a hypoxia-inducible promoter.

example 3

[0418] 6.3 Example 3: Vectors based on hyperglycosylated bevacizumab Fab cDNA

[0419] A hyperglycosylated bevacizumab Fab cDNA-based vector comprising a transgene comprising the bevacizumab Fab of the light and heavy chain cDNA sequences (SEQ ID NO. 10 and 11, respectively) was constructed. A portion having a mutation in the sequence encoding one or more of the following mutations: L118N (heavy chain), E195N (light chain) or Q160N or Q160S (light chain). Transgenes also include nucleic acids comprising a signal peptide selected from the group listed in Table 1. The nucleotide sequences encoding the light and heavy chains are separated by an IRES element or 2A cleavage site to generate a bicistronic vector. Optionally, the vector additionally includes a hypoxia-inducible promoter.

[0420] 6.4 Example 4: Hyperglycosylated ranibizumab cDNA based vector

[0421] A vector based on the hyperglycosylated ranibizumab Fab cDNA comprising the transgene comprising the ranibizumab li...

example 7

[0476] 6.7 Example 7: Open-label Phase 2a Dose Evaluation of Construct II Gene Therapy in Participants with Diabetic Retinopathy

[0477] This example is an updated version of Example 6 and provides an overview of the Phase 2a dose evaluation of Construct II gene therapy in participants with diabetic retinopathy (DR). Continued stable expression of the construct II transgene product following a single gene therapy treatment for DR could potentially reduce the treatment burden of currently available therapies while preserving vision, its favorable benefit:risk spectrum. The current proof-of-concept study was designed to evaluate the safety and efficacy of Construct II gene therapy at 2 different dose levels in participants with DR.

[0478] 6.7.1 Purpose and Endpoints

[0479] Table 6: Primary Purposes and Endpoints and Secondary Purposes and Endpoints

[0480]

[0481]

[0482]

[0483] AAV8 = adeno-associated virus serotype 8; AE = adverse event; CI-DME = centrally...

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