(21) International Application Number

(21) International Application Number

( (51) International Patent Classification: EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, C12N 15/86 (2006.0 1) A61K 9/51 (2006.0 1) MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, C07K 14/705 (2006.01) TR), OAPI (BF, BJ, CF, CG, Cl, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). (21) International Application Number: PCT/US2020/049087 Declarations under Rule 4.17: (22) International Filing Date: — as to the applicant's entitlement to claim the priority of the 02 September 2020 (02.09.2020) earlier application (Rule 4.17(iii)) — of inventorship (Rule 4.17(iv)) (25) Filing Language: English Published: (26) Publication Language: English — with international search report (Art. 21(3)) (30) Priority Data: — with sequence listing part of description (Rule 5.2(a)) 62/895,454 03 September 2019 (03.09.2019) US 63/056,5 14 24 July 2020 (24.07.2020) US (71) Applicant: SANA BIOTECHNOLOGY, INC. [US/US]; 188 East Blaine Street, Suite 400, Seattle, Washington 98102 (US). (72) Inventors: EMMANUEL, Akinola Olumide; 188 East Blaine Street, Suite 400, Seattle, Washington 98102 (US). ENNAJDAOUI, Hanane; 188 East Blaine Street, Suite 400, Seattle, Washington 98102 (US). JAIN, Suvi; 188 East Blaine Street, Suite 400, Seattle, Washington 98102 (US). RUZO MATIAS, Alberto; 188 East Blaine Street, Suite 400, Seattle, Washington 98 102 (US). SHAH, Jagesh V.; 188 East Blaine Street, Suite 400, Seattle, Washington 98102 (US). (74) Agent: POTTER, Karen et al.; Morrison & Foerster LLP, 1253 1 High Bluff Drive, Suite 100, San Diego, California 92130-2040 (US). (81) Designated States (unless otherwise indicated, for every kind of national protection available) : AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, IT, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, WS, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available) : ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, (54) Title: CD24-ASSOCIATED PARTICLES AND RELATED METHODS AND USES THEREOF (57) Abstract: Provided herein are non-cell particles, e.g. virus particles or virus-like particles, such as pseudotyped lentiviral-like particles, containing an exogenous CD24 or a biologically active portion of CD24. In some embodiments, the non-cell particles, e.g. virus particles or virus-like particles, such as pseudotyped lentiviral-like particles, can further contain an exogenous CD47 or a biolog¬ ically active portion of CD47. Also provided herein are compositions containing such non-cell particles and methods of making and using the non-cell particles. CD24-ASSOCIATED PARTICLES AND RELATED METHODS AND USES THEREOF Cross-Reference to Related Applications [0001] This application claims priority to U.S. provisional application 62/895,454, filed September 3, 2020, and to U.S. provisional application 63/056,514, filed July 24, 2020, the contents of each of which are incorporated by reference in their entirety for all purposes. Incorporation by Reference of Sequence Listing [0002] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 186152003740SeqList.TXT, created September 2, 2020, which is 202,892 bytes in size. The information in the electronic format of the Sequence Listing is incorporated by reference in its entirety. Field [0003] The present disclosure relates to non-cell particles, e.g. vims particles or vims-like particles, such as pseudotyped lentiviral-like particles, containing an exogenous CD24 or a biologically active portion of CD24. In some embodiments, the non-cell particles, e.g. vims particles or virus-like particles, such as pseudotyped lentiviral-like particles, can further contain an exogenous CD47 or a biologically active portion of CD47. The present disclosure also provides compositions containing such non-cell particles and methods of making and using the non-cell particles. Background [0004] Non-cell particles, such as lentiviral-like vector particles, are commonly used in gene therapy for delivery of exogenous agents to cells. When administered to subjects, however, such particles can be recognized as foreign and can be engulfed or scavenged by phagocytic cells. Phagocytosis is conducted primarily by highly specialized cells, such as macrophages, monocytes and neutrophils, with the goal of clearing pathogens and/or other foreign invaders that the phagocytic cell recognizes through an array of specialized pattern recognition receptors. Phagocytosis of particles, such as lentiviral vector particles, limits the in vivo half-life and their effectiveness as gene therapy vectors. Thus, there is a long felt need for effective particle delivery vehicles, including lentiviral-like particles along with other types of non-cell particles, for effectively evading the phagocytic cell uptake of the immune system, thereby resulting in a reduction of an immune response. The provided disclosure addresses this need. Summary [0005] Provided herein is a non-cell particle comprising CD24 or a biologically active portion thereof on an exposed surface of the particle, wherein the non-cell particle is 1 pm or smaller. In some embodiments, the CD24 or the biologically active portion thereof binds Siglec- 10. In some embodiments, the CD24 or biologically active portion thereof is human. [0006] In some embodiments, the CD24 or biologically active portion thereof (i) comprises the sequence set forth in SEQ ID NO: 2; (ii) an amino acid sequence having at least at or about 90%, at least at or about 91 , at least at or about 92%, at least at or about 93%, at least at or about 94%, at least at or about 95%, at least at or about 96%, at least at or about 97%, at least at or about 98%, or at least at or about 99% sequence identity to SEQ ID NO:2 that binds Siglec- 10; or (iii) a binding portion of (i) or (ii) that binds to Siglec-10. In some embodiments, the CD24 or biologically active portion thereof comprises the sequence set forth in SEQ ID NO:2. [0007] In some embodiments, the CD24 or biologically active portion is displayed on an exposed surface of the particle via a Glycosylphosphatidylinositol (GPI) membrane anchor. [0008] In some embodiments, the CD24 or the biologically active portion thereof is encoded by a nucleic molecule encoding the sequence set forth in SEQ ID NOG or an amino acid sequence having at least at or about 90%, at least at or about 91%, at least at or about 92%, at least at or about 93%, at least at or about 94%, at least at or about 95%, at or about 96%, at least at or about 97%, at least at or about 98%, or at least at or about 99% sequence identity to SEQ ID NOG that binds Siglec-10. In some embodiments, the CD24 or the biologically active portion thereof is encoded by a nucleic molecule encoding the sequence set forth in SEQ ID NO: 15 or an amino acid sequence having at least at or about 90%, at least at or about 91%, at least at or about 92%, at least at or about 93%, at least at or about 94%, at least at or about 95%, at or about 96%, at least at or about 97%, at least at or about 98%, or at least at or about 99% sequence identity to SEQ ID NO: 15 that binds Siglec-10. [0009] In some embodiments, the CD24 or biologically active portion is displayed on an exposed surface of the particle via a transmembrane domain. [0010] In some embodiments, the CD24 or biologically active portion is a glycoprotein. In some embodiments, the CD24 or biologically active portion is sialylated. In some embodiments, the CD24 or biologically active portion comprises an a2-3-linked sialoside and/or an a2-6- linked sialoside. In some embodiments, the CD24 or biologically active portion has a molecular weight of between at or about 35 kDa and at or about 45 kDa. [0011] In some embodiments, the non-cell particle further comprises a CD47 or a biologically active portion thereof on an exposed surface of the non-cell particle. In some embodiments, the CD47 or biologically active portion binds to SIRPa. In some embodiments, the CD47 or biologically active portion is human. In some embodiments, the CD47 or biologically active portion comprises the extracellular domain or CD47 or a binding portion thereof that binds to SIRPa. [0012] In some embodiments, the CD47 or biologically active portion thereof (i) comprises the sequence set forth in SEQ ID NO: 7; (ii) an amino acid sequence having at least at or about 90%, at least at or about 91 , at least at or about 92%, at least at or about 93%, at least at or about 94%, at least at or about 95%, at least at or about 96%, at least at or about 97%, at least at or about 98%, or at least at or about 99% sequence identity to SEQ ID NO:7 that binds to SIRPa; or (iii) a binding portion of (i) or (ii) that binds to SIRPa.

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