Materials and methods for multidirectional biotransportation in virotherapeutics
Provided herein are multispecific molecules comprising a first binding domain that specifically binds to polymeric immunoglobulin receptor (pIgR) and a second binding domain that specifically binds to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and related methods for the treatment of patients infected with SARS-CoV-2.
1. A multispecific molecule comprising: (a) a first binding domain that specifically binds to polymeric immunoglobulin receptor (pIgR), and (b) a second binding domain that specifically binds to SARS-COV-2, wherein the first binding domain comprises a single-domain antibody (VHH) comprising:
(a) a CDR1 comprising the amino acid sequence of SEQ ID NO:1, a CDR2 comprising the amino acid sequence of SEQ ID NO:2, and a CDR3 comprising the amino acid sequence of SEQ ID NO:3;
(b) a CDR1 comprising the amino acid sequence of SEQ ID NO:4, a CDR2 comprising the amino acid sequence of SEQ ID NO:5, and a CDR3 comprising the amino acid sequence of SEQ ID NO:6;
(c) a CDR1 comprising the amino acid sequence of SEQ ID NO:7, a CDR2 comprising the amino acid sequence of SEQ ID NO:8, and a CDR3 comprising the amino acid sequence of SEQ ID NO:9;
(d) a CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a CDR2 comprising the amino acid sequence of SEQ ID NO: 11, and a CDR3 comprising the amino acid sequence of SEQ ID NO:12;
(e) a CDR1 comprising the amino acid sequence of SEQ ID NO:13, a CDR2 comprising the amino acid sequence of SEQ ID NO:14, and a CDR3 comprising the amino acid sequence of SEQ ID NO:15;
(f) a CDR1 comprising the amino acid sequence of SEQ ID NO:17, a CDR2 comprising the amino acid sequence of SEQ ID NO: 18, and a CDR3 comprising the amino acid sequence of SEQ ID NO:19;
(g) a CDR1 comprising the amino acid sequence of SEQ ID NO:20, a CDR2 comprising the amino acid sequence of SEQ ID NO:21, and a CDR3 comprising the amino acid sequence of SEQ ID NO:22;
(h) a CDR1 comprising the amino acid sequence of SEQ ID NO:23, a CDR2 comprising the amino acid sequence of SEQ ID NO:24, and a CDR3 comprising the amino acid sequence of SEQ ID NO:25;
(i) a CDR1 comprising the amino acid sequence of SEQ ID NO:26, a CDR2 comprising the amino acid sequence of SEQ ID NO:27, and a CDR3 comprising the amino acid sequence of SEQ ID NO:28; or
(j) a CDR1 comprising the amino acid sequence of SEQ ID NO:29, a CDR2 comprising the amino acid sequence of SEQ ID NO:30, and a CDR3 comprising the amino acid sequence of SEQ ID NO:31;
and wherein the second binding domain comprises a peptide comprising:
(a) angiotensin-converting enzyme 2 (ACE2);
comprising the amino acid sequence of SEQ ID NO: 194;
(b) the extracellular domain of ACE2;
comprising the amino acid sequence of SEQ ID NO: 134; or
(c) a truncated extracellular domain of ACE2;
comprising the amino acid sequence of SEQ ID NO: 120 or the amino acid sequence of SEQ ID NO: 121.
2. The molecule of claim 1 , wherein the molecule is a bispecific molecule.
3. The molecule of claim 1 , wherein the second binding domain specifically binds to the surface of SARS-COV-2.
4. The molecule of claim 1 , wherein the first binding domain comprises a VHH comprising:
(a) a complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO:1, a CDR2 comprising the amino acid sequence of SEQ ID NO:2, and a CDR3 comprising the amino acid sequence of SEQ ID NO:3; further wherein the VHH comprises the amino acid sequence of SEQ ID NO:16; or
(b) a CDR1 comprising the amino acid sequence of SEQ ID NO:17, a CDR2 comprising the amino acid sequence of SEQ ID NO:18, and a CDR3 comprising the amino acid sequence of SEQ ID NO:19; further wherein the VHH comprises the amino acid sequence of SEQ ID NO:32.
5. The molecule of claim 1 , wherein the first binding domain specifically binds to pIgR that is present on the mucosal endothelium.
6. The molecule of claim 1 , wherein the SARS-COV-2 is neutralized when the molecule specifically binds to the pIgR and to SARS-COV-2.
7. A nucleic acid encoding the molecule of claim 1 .
8. A vector comprising the nucleic acid of claim 7 .
9. A host cell comprising the vector of claim 8 .
10. A kit comprising the vector of claim 8 and packaging.
11. A pharmaceutical composition comprising the molecule of claim 1 , and a pharmaceutically acceptable carrier.
12. A method of producing a pharmaceutical composition comprising combining the molecule of claim 1 with a pharmaceutically acceptable carrier to obtain the pharmaceutical composition.
13. A method of inhibiting host cell entry or proliferation of target cells expressing SARS-COV-2, the method comprising contacting the target cells with the molecule of claim 1 , wherein contacting the target cells with the molecule inhibits host cell entry or proliferation of the target cells.
14. A method of treating SARS-COV-2 or COVID-19 in a subject, comprising administering an effective amount of the molecule of claim 1 to the subject.
15. The molecule of claim 3 , wherein the second binding domain specifically binds to the spike glycoprotein on the surface of SARS-COV-2.
16. The molecule of claim 3 , wherein the second binding domain specifically binds to the S1 subunit of the spike glycoprotein on the surface of SARS-COV-2.
17. The molecule of claim 5 , wherein the first binding domain specifically binds to pIgR that is present on the lung mucosal endothelium.
18. The molecule of claim 6 , wherein SARS-COV-2 is neutralized with an EC 50 of (a) less than about 4 nM; (b) less than about 3 nM; (c) less than about 1 nM; (d) less than about 500 pM; or (e) less than about 100 pM.