Engineered platelets for targeted delivery of therapeutic agents

Abstract

The present invention provides engineered platelets having a chimeric platelet receptor (CPR) with desired target specificity. Furthermore, the engineered platelets may contain cargo that can be released upon platelet activation. In addition, the platelets can be generated in vitro from megakaryocytes engineered to produce non-thrombotic platelets. [Solution] To provide an engineered megakaryocyte or precursor capable of reducing thrombogenic ability and / or producing platelets with reduced thrombogenic ability, comprising the disruption or deletion of at least one gene encoding a protein involved in the recognition of a primary stimulus for thrombosis, one gene encoding a protein involved in the recognition of a secondary mediator for thrombosis, and one gene encoding a protein involved in the release of a secondary mediator for thrombosis.

Description

[Technical Field]

[0001] Sequence List This application is filed together with an electronic sequence listing. The sequence listing file, titled SEQLIST.txt, was created on December 14, 2020, and has a size of 46,300 bytes. The electronic information of the sequence listing is incorporated herein by reference in its entirety.

[0002] The present invention relates to engineered platelets for targeted therapeutic delivery systems. [Background technology]

[0003] Platelets are small, enucleated cells that cannot divide or replicate. In the human body, platelets play a vital role in recognizing damaged tissue and releasing their contents to reduce or prevent bleeding. Thrombopoietin from the kidneys and liver comes into contact with bone marrow stem cells, triggering differentiation into megakaryoblasts, and additional signaling leads to differentiation of megakaryoblasts into megakaryocytes, the precursors of megakaryoblasts. Megakaryocytes are large cells with elongated platelet precursors that germinate fragments as they divide and proliferate, producing platelets.

[0004] Mitochondria, microtubules, and vesicles are contained within platelets. Platelets have a lifespan of approximately 10 days and are cleared by macrophages. Platelets are approximately 7 μm in size. 3 Platelets have a volume and a diameter of 300 nm. Platelets are metabolically active and can alter gene expression through post-transcriptional regulation of preloaded mRNA expression (e.g., by miRNA). Upon activation, granulation is stimulated, changing their shape and releasing the contents of intracellular vesicles.

[0005] Platelets respond to a variety of extracellular signals through a diverse set of signaling pathway receptors. These receptors act both to trigger intracellular signaling cascades that result in platelet degranulation and effector release, and to induce platelet aggregation and adhesion. Glycoprotein VI platelet (GPVI) signaling functions similarly to many immune cell receptors, such as TCRs. Interestingly, platelets also express Toll-like receptors (TLRs), which can mediate targeted bacterial death through peptide secretion and immune system activation.

[0006] Regarding platelet degranulation, a wide variety of products are released. Vesicles are released by exocytosis. Platelets contain three main subtypes of vesicles: α-granules (50-80 per platelet), dense granules (3-8 per platelet), and large dense-core vesicles (LDCVs) (approximately 10,000 per platelet). Different mutations can selectively disrupt the biosynthesis of each vesicle subtype.

[0007] Alpha-granules have a diameter of approximately 200-500 nm and account for about 10% of the platelet's volume. Most effector proteins are found in alpha-granules. For example, effector proteins released from α-granules include intrinsic membrane proteins such as P-selectin, αIIbβ, and GPIbα; coagulants / anticoagulants and fibrinolytic proteins such as factor V, factor IX, and plasminogen; adhesion proteins such as fibrinogen and von Willebrand factor (vWF); chemokines such as CXCL4 (cytokine (CXC motif) ligand 4), also known as platelet factor 4 or PF4, and CXCL12 (cytokine (CXC motif) ligand 12), also known as stromal cell-derived factor 1 alpha or SDF-1α; growth factors such as elongation growth factor (EGF) and insulin-like growth factor 1 (IGF); angiogenic factors / inhibitors such as vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and angiostatins; and immune mediators such as immunoglobulin G (IgG) and complement precursors.

[0008] Dense granules have a diameter of approximately 150 nm and account for about 1% of the platelet volume. Effector proteins released from dense granules include Ca 2+ and Mg 2+ Examples include cations such as ion, polyphosphates, physiologically active amines such as serotonin and histamine, and nucleotides such as adenosine diphosphate (ADP) and adenosine triphosphate (ATP).

[0009] LDCVs have a diameter ranging from approximately 150 nm to 300 nm and account for about 13.5% of the platelet volume. Effector proteins released from LDCVs include structural proteins (e.g., granin and glycoproteins), vascoregulators (e.g., cateholamine, vasostatin, renin-angiotensin), paracrine signaling factors (e.g., guanylin, neurotensin, chromogranin B), immune mediators (e.g., enkelytin and ubiquitin), opioids (e.g., enkephalin and endorphins), and ions (e.g., Ca 2+ Examples include nucleotides (Na+, Cl-), as well as nucleotides and polyphosphates (e.g., adenosine monophosphate (AMP), guanosine diphosphate (GDP), uridine-5'-triphosphate (UTP)). [Overview of the Initiative] [Problems that the invention aims to solve]

[0010] Current cell therapies based on engineered chimeric antigen receptor T cells (CAR-T cells) have shown promise in treating cancer, but their use is limited to a small number of patients due to safety concerns, particularly regarding oncogenic transformation in patients, and the limited ability to produce generic or universal therapeutic products. There is a long-recognized need in this field for a new type of therapy that has the potential to treat cancer, autoimmune conditions, and infections, free from the safety, cost, and patient matching issues that plague current cell therapy products. [Means for solving the problem]

[0011] Various embodiments of the present invention described herein are a) A platelet-stimulating domain, comprising an intracellular domain including a domain from the immune receptor tyrosine-based activation motif (ITAM) receptor, b) Provide a chimeric platelet receptor (CPR) comprising a heterologous targeting domain that recognizes and binds to a target.

[0012] A heterologous targeting domain means that the targeting domain is heterologous to the intracellular platelet-stimulating domain; in other words, the targeting domain is not a typical extracellular domain associated with an intracellular domain. A heterologous targeting domain can bind to endogenous targets, for example, to tumor antigens that are endogenous to the target, but because CPR is a chimeric molecule, the targeting domain is heterologous to the intracellular platelet-stimulating domain.

[0013] The present invention as described herein also, A first region encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 1-19, 24-47, and 52-55; (i) a linker or targeting domain encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 48-51; and (ii) myelin oligodendrocyte glycoprotein (MOG), glutamate decarboxylase 2 (GAD65), myelin-associated glycoprotein (MAG), peripheral myelin protein 22 (PMP22), thyroid peroxidase (TPO), voltage-opening potassium channel (VGKC), proteolipid protein (PLP), acetylcholine receptor (AChR), tribulous pseudokinase 2 ( TRIB2), N-methyl-D-aspartate (NMDA) type glutamate receptor (GluR), glutamate decarboxylase 2 (GAD2), armadillo repeat 9 (ARMC9), cytochrome P450 family 21 subfamily A member 2 (CYP21A2), calcium-sensing receptor (CASR), nuclear autoantigenic sperm protein (NASP), insulin, thyroid-stimulating hormone receptor (TSHR), tyloperoxidase, acioglycoprotein receptor, cytochrome P450 family 2 subfamily D member 6 (CYP2D6), lactoferrin (LF), tissue transglutaminase (TTG), H / KATP-ase, Factor XIII (F8), Beta-2-glycoprotein I (Beta-2-GPI), Erythrocyte I / I, B2 integrin (ITGB2), Granulocyte colony-stimulating factor (G-CSF), Glycoprotein (GP) IIb / IIa, Collagen II (COLII), Fibrinogen (FBG) βα, Myeloperoxidase (MPO), Cardiac myosin (CYO), Proteinase 3 (PRTN3), Trichohyalin (TCHH), Bullous pemphigoid-related (BP), Glycoprotein 1 (GP1), Laminin-332 (LM332), Transglutaminase (TGM), Collagen VII (COLVII), P80 Coilin (COIL), Desmoglein I (DSG1), Desmoglein III (DSG3), SRY-Box 10 (SOX1) (0) At least a portion of proteins selected from the group consisting of nuclear small molecular weight ribonucleoprotein U1 subunit (70SNRNP70), S antigen (SAG), and collagen alpha-3(IV) chain (α3(IV)NC1 collagen), and (iii) 3F8, 8H9, avagovomab, absiximab, abituzumab, abrezekimab, abrilumab, actoxumab, adalimumab, adecatumumab, atidortoxumab, aducanumab, afasevikumab, afelimomab, alacizumab pego pego), alemtuzumab, alirocumab, altumomab pentetate, amatuximab, anatumomab mafenatox, anddecaliximab, anetumab ravtansine, aniflorumab, anlukinzumab, apolizumab, aprutumab ixadotinixadotin), alsitumomab, ascrinvacumab, aselizumab, atezolizumab, atinumab, atorolimumab, avelumab, azintuxizumab vedotin Vedotin, Bapineuzumab, Basiliximab, Babituximab, BCD-100, Bectumomab, Begelomab, Belantamab-Fodotin, Berimumab, Bemarituzumab, Benralizumab, Berlimatoxumab, Bermekimab, Bersanlimab, Bertilimumab, Besilesomab, Bevacizumab, Bezlotoxumab, Biciromab Bimagrumab, bimekizumab, virtamimab, vibatuzumab meltansine, preserumab, blinatumomab, blontuvetmab, brosozumab, vococizumab, brazikumab, brentuximab vedotin, briakinumab, brodalumab, brolucizumab, bronticutuzumab, brosumab, kabilizumab, camidanrumab tesirin, camrelizumab, canakinumab, cantuzumab meltansine, cantuzumab brabutansine ravtansine), caplacizumab, capromab pendetide, carlumab, carotuximab, catumakisomab, cbr96-doxorubicin immune complex, cedelizumab, semiprimab, cergutuzumab amnaleukin, certolizumab pegol, cetrelimab, cetuximab, cibisatamab, cirmtuzumab, citatuzumab vodatoxbogatox), cixutumumab, crazakizumab, clenoliximab, cribatuzumab tetraxetan, codrituzumab, cofetuzumab peridotin, coltuximab labutancin ravtansine), conatumumab, concizumab, cosflobiximab, crenezumab, chryzanlizumab, clotedumab, CR6261, kusatuzumab, dacetuzumab, dacrizumab, dalotuzumab, dapirolizumab pegol, daratumumab, dectrekumab, demcizumab, denintuzumab mahodotin, denosumab, depatuxizumab mahodotin, derlotuximab biotin biotin), detumomab, dezamizumab, dinutuximab, diridavumab, domagrozumab, Dorlimomab aritox, dostallimab, drozitumab, DS-8201, duligotuzumab, dupilumab, durvalumab, dusigitumab, duvoltuxizumab, ecromeximab, eculizumab, edobacomab, edrecolomab, efalizumab, efungumab Eldelumab, Elezanumab, Elgemtumab, Elotuzumab, Elsilimomab, Emactuzumab, Emapalmab, Emibetuzumab, Emicizumab, Enapotamab Vedotin, Enavatuzumab, Enfortumab Vedotin, Enlimomab Pegolpegol), enoblituzumab, enokizumab, enoticumab, ensituximab, epitumomabcituxetan), epratuzumab, eptinezumab, erenumab, erzmaxomab, etaracizumab, etigilimab, etrolizumab, evonacumab, evolocumab, exvivirumab, fanolesomab, faralimomab, falisimab, falletuzumab, facinumab numab), FBTA05, Felvizumab, Fezakinumab, Fibatuzumab, Ficlatuzumab, Figitumumab, Firivumab, Flambotumab, Fletikumab, Flotetuzumab, Fortrizumab, Foralumab, Foravirumab, Fremanezumab, Fresolimmumab (Fr Esolimumab, flubokimab, flunevetmab, fluranumab, futuximab, galcanezumab, galiximab, gancotamab, ganitumab, gantenerumab, gatipotuzumab, gavilimomab, gedivumab, gemtuzumab ozogamicin, gebokizumab, gilvetmab, jimcirma Gimsilumab, Girentuximab, Grembatumumab Vedotin, Golimumab, Gomiliximab, Gosuranemab, Guselkumab, Ianalumab, Ibalizumab, IBI308, Ibritumomab Chiuxetan, Ikurumab, Idarucizumab, Ifabotuzumab, Igovomab, Iladatuzumab VedotinIndusatumab vedotin, IMAB362, Imalumab, Imaprelimab, Imisilomab, Imgatuzumab, Inclacumab, Indatuximab ravtansine, Indusatumab vedotin Vedotin, Inebilizumab, Infliximab, Intetumumab, Inolimomab, Inotuzumab Ozogamicin, Ipilimumab, Iomab-B, Iratumumab, Isatuximab, Iscarimab, Istilatumab, Itorizumab, Ixekizumab, Keliximab, Labetuzumab, Lacnotuzumab, Radilatuzumab Vedotin, Lampalizumab, Lanadelumab, Landogrozumab, Laprituximab Emtansine emtansine), Larcabiximab, Lebrikizumab, Lemalesomab, Lendalizumab, Lenvervimab, Lenzilumab, Lerdelimumab, Leronlimab, Lesofavumab, Letolizumab, Lexatumumab, Libivirumab, Rifastuzumab Vedotin, Rigerizumab, Ronkastuzumab Tecilin, Rosatuzumab Vedotin xizumab vedotin), lilotomab satetraxetan, lintuzumab, lirilumab, lodelcizumab, lokivetmab, rorbotuzumab meltansine, lucatumumab, rulizumab pegol, lumiliximab, rumretuzumab, rupatumab amadotin amadotin), rituximab, mapatumumab, margetuximab, marstacimab, maslimomab, mavrilimumab, matsuzumab, mepolizumab, metelimumab, milatuzumab, minretumomab, mirikizumab umab), milbetuximab sorabtansine, mitumomab, modotuximab, mogamulizumab, monalizumab, morolimumab, mosunetuzumab, motavizumab, moxetumomab pasudotox, muromonab-CD3, nacolomab fenatox Tafenatox), Namilumab, Naptumomab Estafenatox, Naratuximab emtansine, Narnatumab, Natalizumab, Navixixizumab, Navivumab, Naxitamab, Nebacumab, Necitumumab, Nemolizumab, NEOD001, Nerelimomab, Nesvacumab, Netakimab, Nimotuzumab, Nilsevimab, Nivolumab, Nofetumomab MerpentanMerpentan), Obiltoxaximab, Obinutuzumab, Ocaratuzumab, Ocrelizumab, Odulimomab, Ofatumumab, Olaratumab, Olekrumab, Olendalizumab, Olokizumab, Omalizumab, O Omburtamab, OMS721, Onartuzumab, Ontuxizumab, Onvatilimab, Opicinumab, Opotuzumab Monatox, Oregovomab, Orticumab, Otelixizumab Otiliumab, Otlertuzumab, Oxelumab, Ozanezumab, Ozoralizumab, Pagibaximab, Palivizumab, Pamrevlumab, Panitumumab, Pankomab, Panobacumab, Parsatuzumab, Pascoli Pascolizumab, Pasotuxizumab, Pateclizumab, Patritumab, PDR001, Pembrolizumab, Pemtumomab, Perakizumab, Pertuzumab, Pexelizumab, Pizilizumab, Pinatuzumab VedotinVedotin, Pintumomab, Placulumab, Prozalizumab, Pogalizumab, Polatuzumab Vedotin, Ponezumab, Porgaviximab, Placinezumab, Prezalizumab, Priliximab, Pritoxaximab, Pritumumab, PRO140, Quilizumab, Racotumomab, Radret umab), rafivirumab, ralpancizumab, ramucirumab, ranevetmab, ranibizumab, laxibakumab, lavagalimab, ravulizumab, refanezumab, regavirumab, relatlimab, remtolumab, reslizumab, rilotumumab, rinucumab, risankizumab, rituximab, rivabazumab pegol pegol), Robatumumab, Rmab, Rolledumab, Romilkimab, Romosozumab, Rontalizumab, Rosmantuzumab, Robalpituzumab Tecilin, Robelizumab, Rozanolixizumab, Ruplizumab, SA237, Sacituzumab Govitecan, Samalizumab, Samrotamab VetotinVedotin, sarilumab, satralizumab, satumomab pendetide, secukinumab, selicrelumab, seribantumab, setoxaximab, setrusumab, sevilumab, sibrotuzumab, SGN-CD19A, SHP647, sifalimumab, siltuximab, simtuzumab, siplizumab, siltratumab vetotin Vedotin, silukumab, sofituzumab, solanezumab, solitomab, sonepcizumab, sontuzumab, spartalizumab, stamulumab, sulesomab, suptavumab, sutimlimab, suvizumab, subutiratoxumab, tabalumab, tetraxetan (tacatuzumab) tetraxetan), tadocizumab, talacotuzumab, talizumab, tamtuvetmab, tanezumab, taplitumomab paptox, tarextumab, tavolimab, tefibazumab, telimomabaritox), terisotuzumab vedotin, tenatumomab, teneliximab, teplizumab, tepoditamab, teprotumumab, tesidolumab, tetulomab, tezepelumab, TGN1412, tibulizumab, tildrakizumab, tigatuzumab, timigutuzumab, timolumab, tiragolumab ab), Tislelizumab, Tisotumab Vedotin, TNX-650, Tocilizumab, Tomzotuximab, Trallizumab, Tosatoxumab, Toshitumomab, Tovetumab, Tralokinumab, Trastuzumab, Trastuzumab Emtansine, TRBS07, Tregalizumab, Tremelimumab, Trevogrumab, Tucotuzumab Cermoloukin celmoleukin), tuvirumab, ublituximab, urocuplumab, urelumab, urtoxazumab, ustekinumab, utomilumab, vadastuximab talirine, vanalimab, polatuzumab vedotinVedotin, Vantictumab, Vanucizumab, Vapaliximab, Varisacumab, Varilumab, Vatelizumab, Vedolizumab, Veltuzumab, Vepalimomab, Vesencumab, Visilizumab, Vobarilizumab, Voloximab, Vonlerolizumab, Vopratelimab, Borsetuzumab mahodotin, Votumumab, Vunakizumab, Xentuzumab, XMAB-5574, Zalutumumab, Zanolimumab, Zatuximab, Zenocutuzumab, Ziralimumab, Zolbetuximab, Zolimomab aritox The present invention provides a chimeric platelet receptor (CPR) comprising an antibody or antibody fragment selected from the group consisting of (iv) aritox, and a second region encoded by a nucleic acid sequence selected from the group consisting of (iv) a major histocompatibility complex (MHC) class 1 receptor or a major histocompatibility complex (MHC) class 2 receptor, wherein the MHC class 1 receptor binds to peptides derived from tumor antigens, novel antigens, or autoantigens, and the MHC class 2 receptor binds to peptides derived from tumor antigens, novel antigens, or autoantigens.

[0014] In some embodiments, the chimeric platelet receptor binds to at least one antigen. In some embodiments, the chimeric platelet receptor binds to tissues in the body of the subject. In some embodiments, the chimeric platelet receptor inhibits platelet activation. In some embodiments, the chimeric platelet receptor is an ITAM-containing receptor. In some embodiments, the chimeric platelet receptor stimulates platelet activation. In some embodiments, the chimeric platelet receptor is an ITAM-containing receptor. In some embodiments, the chimeric platelet receptor binds to at least one antigen that is an endogenous antigen, for example, an endogenous host or target antigen. The endogenous host or target antigen means that the antigen is present in the host or subject to which the chimeric platelet receptor is administered, or to which cells such as megakaryocytes or platelets constituting the CPR of the present invention are administered. For example, the antigen may be an antigen present in a tissue, or a specific subset of a tissue, or in the plasma or blood of a subject, for example, a human subject. The antigen may be an antigen that is expressed at an abnormal level, e.g., an abnormally high level, on tissues that do not normally express high levels of the antigen, or that do not normally express the antigen at all.

[0015] In some embodiments, the chimeric platelet receptor is neither a GPCR nor a protease-activated receptor, nor does it contain them.

[0016] The present invention also provides nucleic acids encoding the chimeric platelet receptor of the present invention. In preferred examples, since the chimeric platelet receptor is not a naturally occurring receptor, the nucleic acid encoding the receptor is not a naturally occurring nucleic acid. In some embodiments, the nucleic acid encoding the CPR of the present invention also comprises heterologous nucleic acid sequences. In some cases, the nucleic acid is operably linked to an expression regulatory sequence. The expression regulatory sequence is thought to include components such as enhancers and promoters. In one embodiment, the nucleic acid of the present invention comprises a heterologous promoter. In the same or different embodiments, the nucleic acid of the present invention comprises a heterologous enhancer sequence.

[0017] In some embodiments, the nucleic acid is DNA. In some embodiments, the nucleic acid is RNA, for example, mRNA. In some embodiments, the nucleic acid comprises a megakaryocyte-specific promoter or a platelet-specific promoter. The terms megakaryocyte-specific promoter and platelet-specific promoter are used synonymously. Those skilled in the art will understand what the terms megakaryocyte-specific promoter and platelet-specific promoter mean.

[0018] The present invention also provides a vector comprising a nucleic acid encoding CPR. The term vector includes plasmids. In some embodiments, the vector also comprises a heterologous nucleic acid. In some embodiments, the vector comprises a megakaryocyte-specific promoter. In some embodiments, the vector comprises a platelet-specific promoter.

[0019] The present invention also provides a viral particle or viral vector comprising any one or more of the nucleic acids of the present invention.

[0020] The present invention also provides a nucleic acid encoding a cargo protein or peptide comprising a sequence suitable for driving expression in megakaryocytes and / or platelets. For example, in some embodiments, the nucleic acid encoding the cargo protein, cargo peptide or cargo RNA is operably linked to a heterologous expression control sequence such as a promoter. In some embodiments, the nucleic acid encodes a cargo protein or peptide and comprises a megakaryocyte-specific promoter or a platelet-specific promoter. In some embodiments, the nucleic acid encodes a cargo protein or peptide and comprises a heterologous sequence such as a megakaryocyte-specific promoter or a platelet-specific promoter.

[0021] Various embodiments of the present invention described herein provide a therapeutic delivery system comprising an engineered platelet presenting the chimeric platelet receptor described above and at least one therapeutic agent selected from the group consisting of a toxin, a protein, a small molecule drug, and a nucleic acid packaged within vesicles in the platelet.

[0022] In some embodiments, the engineered platelets are produced from iPSC precursors. In some embodiments, the nucleic acids are messenger ribonucleic acid (mRNA), microRNA (miRNA), short hairpin RNA (shRNA), and clustered regularly interspaced short palindromic repeat (CRISPR) sequences. In some embodiments, the proteins are selected from the group consisting of antibodies, enzymes, cytokines, and CRISPR-associated protein 9 (Cas9). In one aspect, the enzyme is a nuclease.

[0023] In some embodiments, the nuclease is a transcription activator-like effector nuclease (TALEN). In some embodiments, the antibodies bind to targets such as, but not limited to, tumor antigens or neoantigens. In some embodiments, the therapeutic agent is released from the platelets following activation of the platelets by an antigen recognized by a chimeric platelet receptor.

[0024] Various methods are provided for delivering cargo to a subject that requires the cargo. As described herein, the cargo can be a therapeutic agent or a toxin. The cargo can be a protein or a peptide, or can be a nucleic acid such as a therapeutic RNA or mRNA. The preference for the cargo is as described elsewhere in this specification. The present invention provides a method of delivering cargo, comprising administering an effective amount of any one or more of the engineered megakaryocytes, engineered platelets, and / or CPRs described in any one of the preceding claims. The present invention also provides a therapeutic delivery system. The present invention also provides a non-therapeutic delivery system. The present invention also provides a method for targeted cargo delivery to a target tissue or site in the body, the method comprising administering an effective amount of any one or more of the engineered megakaryocytes, engineered platelets, and / or CPRs described in any one of the preceding claims, wherein the targeting domain of the CPR binds to the target tissue or site in the body.

[0025] Various embodiments of the present invention described herein provide methods for treating a disease, disorder, or condition in a subject, the methods comprising administering the aforementioned therapeutic delivery system to the subject, the chimeric receptor being specific to an antigen associated with the disease, disorder, or condition.

[0026] In some embodiments, the disease, disorder, or condition may be, but is not limited to, cancer, autoimmune, and infectious diseases. In some embodiments, cancer may include acute granulocytic leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, adenocarcinoma, adenosarcoma, adrenal carcinoma, adrenocortical carcinoma, anal carcinoma, anaplastic astrocytoma, angiosarcoma, appendiceal carcinoma, astrocytoma, basal cell carcinoma, B-cell lymphoma), cholangiocarcinoma, bladder carcinoma, bone carcinoma, intestinal carcinoma, brain carcinoma, brainstem glioma, brain tumor, breast carcinoid tumor, cervical carcinoma, cholangiocarcinoma, chondrosarcoma, and chronic cancer. Lymphocytic leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuse astrocytoma, ductal carcinoma in situ, endometrial cancer, ependymoma, epithelioid sarcoma, esophageal cancer, Ewing's sarcoma, extrahepatic cholangiocarcinoma, eye cancer, fallopian tube cancer, fibrosarcoma, gallbladder cancer, stomach cancer, gastrointestinal cancer, gastrointestinal carcinoid cancer, gastrointestinal stromal tumor, systemic, germ cell tumor, glioblastoma pleomorphis, nerve Glioma, pilocytic cell leukemia, head and neck cancer, hemangioendothelioma, Hodgkin's lymphoma, Hodgkin's disease, Hodgkin's lymphoma, hypopharyngeal cancer, invasive ductal carcinoma, invasive lobular carcinoma, inflammatory breast cancer, colon cancer, intrahepatic cholangiocarcinoma, invasive / invasive breast cancer, islet cell carcinoma, jaw cancer, Kaposi's sarcoma, kidney cancer, laryngeal cancer, leiomyosarcoma, leukemia, lip cancer, liposarcoma, liver cancer, lobular carcinoma in situ, low-grade astrocytoma, lung cancer Cancer, lymph node cancer, lymphoma, male breast cancer, medullary carcinoma, medulloblastoma, melanoma, meningioma, Merkel cell carcinoma, mesenchymal chondrosarcoma, mesenchymal mesothelioma, metastatic breast cancer, metastatic melanoma, metastatic squamous cell carcinoma of the neck, mixed glioma, oral cancer, mucinous carcinoma, mucosal melanoma, multiple myeloma, nasal cavity cancer, nasopharyngeal cancer, neck cancer, neuroblastoma, neuroendocrine tumor, non-Hodgkin lymphoma, non-Hodgkin's lymphoma, non-small cell lung cancer, ocular cancer, ocular melanoma, oligodendroglioma, oral cancer, oral cavity cancerCancer, oropharyngeal cancer, osteogenic sarcoma, osteosarcoma, ovarian cancer, ovarian epithelial carcinoma, ovarian germ cell tumor, primary ovarian peritoneal cancer, ovarian cord-stromal tumor, Paget's disease, pancreatic cancer, papillary carcinoma, paranasal sinus cancer, parathyroid cancer, pelvic cancer, penile cancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pilocytic cell astrocytoma, pineal gland tumor, pineoblastoma, pituitary cancer, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, renal pelvis cancer, rhabdomyosarcoma, salivary gland cancer, sarcoma, bone sarcoma, soft tissue sarcoma, uterine sinus cancer, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, spinal cord cancer, spinal column cancer, spinal cord cancer (Spinal cord The group is selected from cancers, spinal tumors, squamous cell carcinomas, gastric cancers, synovial sarcomas, T-cell lymphomas, testicular cancers, throat cancers, thymomas / thymic carcinomas, thyroid cancers, tongue cancers, tonsil cancers, metastatic cell carcinomas, metastatic cell carcinomas, triple-negative breast cancers, fallopian tube cancers (tubal cancers), tubular carcinomas, ureteral cancers, urethral cancers, urethral cancers, uterine adenocarcinomas, uterine cancers, uterine sarcomas, vaginal cancers, and vulvar cancers.

[0027] In some embodiments, the method further comprises incubating the manipulated platelets with at least one therapeutic agent, such as but not limited to toxins, proteins, and small molecule drugs, to generate a therapeutic delivery system. In some embodiments, the nucleic acid may be, but is not limited to, mRNA, miRNA, shRNA, and clustered regularly spaced short palindromic repeat (CRISPR) sequences. In some embodiments, the protein may be, but is not limited to, antibodies, enzymes, and CRISPR-associated protein 9 (Cas9). In some embodiments, the enzyme is a nuclease. In some embodiments, the nuclease is a transcriptional activator-like effector nuclease (TALEN). In some embodiments, incubation occurs before administration.In some embodiments, the disease, disorder, or condition is autoimmune disseminated encephalomyelitis, autoimmune inner ear disease, Batten disease / neuroceroid lipofuscinosis, chronic inflammatory demyelinating polyneuropathy, ophthalmosal encephalitis, anti-basal ganglia, Guillain-Barré syndrome, Hashimoto's encephalopathy, anti-TPO, Isaac syndrome / acquired neurogenic myotonia, Miller-Fischer syndrome, Morvan syndrome, multiple sclerosis, gravitational myasthenia gravis, narcolepsy-pandus, Rasmussen encephalitis, generalized rigidity syndrome, Vogt-Koyanagi-H Arada disease, Addison's disease, autoimmune hypoparathyroidism, autoimmune hypophysitis, autoimmune oophoritis, autoimmune orchitis, polyglandular autoimmune syndrome I (APECED), polyglandular autoimmune syndrome II, polyglandular autoimmune syndrome III, diabetes mellitus, type 1, Graves' disease, Hashimoto's autoimmune thyroiditis, immunodysregulation, polyglandular endocrine disorders, intestinal diseases, X-linked, autoimmune hepatitis type 1, autoimmune hepatitis type 2, autoimmune pancreatitis, abdominal diseases, Crohn's disease, pernicious anemia / atrophic gastritis, primary biliary cirrhosis, primary Sclerosing cholangitis, ulcerative colitis, acquired hemophilia A, antiphospholipid syndrome, autoimmune hemolytic anemia, autoimmune lymphoproliferative syndrome, autoimmune neutropenia, Evans syndrome, Felty syndrome, immune thrombocytopenic purpura, polymyositis / dermatomyositis, relapsing polychondritis, rheumatoid arthritis, Still's disease, alopecia areata, bullous pemphigoid, scarring pemphigoid, herpetiform dermatitis, discoid lupus erythematosus, acquired epidermolysis bullosa, linear scleroderma, pemphigus foliaceus, pemphigus vulgaris, vitiligo, Behçet's disease, Autoimmune diseases such as Churg-Strauss syndrome, Cogan syndrome, Crest syndrome, antifibrillarin, essential mixed cryoglobulinemia, mixed connective tissue disease, POEMS syndrome, scleroderma, Sjögren's syndrome, systemic lupus erythematosus, erythema elevata, Kawasaki disease, microscopic polyangiitis, polyarteritis nodosa, rheumatic fever, Takayasu's arteritis, Wegener's granulomatosis, HLA-B27-associated acute anterior uveitis, sympathetic ophthalmitis, and Goodpasture disease are among the most common.

[0028] The present invention also provides non-therapeutic methods for delivering cargo to subjects that need or desire it. For example, the present invention provides non-therapeutic methods for delivering cargo to subjects that need it. In such non-therapeutic methods, the cargo may be cosmetics. In some embodiments, the present invention provides non-therapeutic methods for targeted delivery of cargo to subjects that need it.

[0029] Various embodiments of the present invention described herein provide engineered platelets produced from megakaryocytes that, compared to platelets produced from unmutated megakaryocytes, include mutations in nucleic acid sequences resulting in disruption of the vesicle biosynthesis pathway or vesicle release pathway in platelets, expression of toxins, or expression of proteins, peptides, or RNA, such as mRNA, e.g., cargo, e.g., therapeutic agents or contrast agents, or deletion of platelet receptors, mediators, or signaling proteins.

[0030] In some embodiments, megakaryocytes are differentiated from iPSC precursors, or megakaryocytes are immortalized. In some embodiments, mutations occur in genes encoding components of the vesicle biosynthesis or vesicle release pathway of the engineered platelets, including but not limited to α-granules, dense granules, and large dense-core vesicles. In some embodiments, the mutation is an insertion into a gene encoding a major histocompatibility complex (MHC) molecule.In some embodiments, the deletions include RAB27a (RAS oncogene), HPS (haptoglobin) gene, integrin AIIbB3, GP1b-IX-V (glycoprotein Ib complexed with glycoprotein IX), Par1 (protease-activated receptor 1), Par4 (protease-activated receptor 4), P2Y1 (purine receptor P2Y1), P2Y12 (purine receptor P2Y12), IP (PGI2R or prostaglandin I2 receptor), and TP (TxA2). R or thromboxane A2 receptor), TLR (Tall-like receptor), GPVI, a2B1 (type 1 collagen receptor), GPIIbIIIA (glycoprotein IIb platelet subunit alpha), CLEC-2 (type C lectin-like receptor 2), MyD88 (myelin differentiation primary response 88), Galphaq (G protein alpha pathway q), LIMK1 (LIM domain kinase 1), vWF (von Willebrand), fibrinogen, PDGF (platelet-derived growth factor), VEGF (Vascular Endothelial Growth Factor), Factor V, Factor VIII, Factor XI, Factor XIII, PF4 (Platelet Factor 4), NAP2 (Nucleosome Assembly Protein 2), Prothrombin, High Molecular Weight Kininogen, Plasminogen Activator Inhibitor 1, α2-Antiplasmin, Plasminogen, P-Selectin, CXCL4 (CXC Motif Chemokine Ligand 4), CXCL7 (CXC Motif Chemokine Ligand 7), FGF (Fibroblast Growth Factor) This includes, but is not limited to, at least one gene such as, EGF (elongation growth factor), HGF (hepatocyte growth factor), IGF (insulin-like growth factor), angiopoietin, thromboxane synthase, PAF (platelet-activating factor), cPLA2a, thrommosponin, CD40L, SgIII (secretogranin III), endostatin, TGF-β (transforming growth factor beta), talin 1, kindlin, and anoctamin 6.

[0031] In some embodiments, the mutation is a deletion, which is a knockout of a gene encoding a prothrombus factor. In some embodiments, the gene is the β2 microglobulin gene, and the deletion results in endogenous MHC class 1 disruption and the production of non-immunogenic platelets. In some embodiments, the mutation reduces the thrombus-forming ability of the engineered platelets compared to platelets produced from unmutated megakaryocytes.

[0032] Various embodiments of the present invention described herein provide a method for reducing activity in the immune system of a subject, the method comprising administering to a subject engineered platelets that display at least one receptor expressing a major histocompatibility complex (MHC) molecule bound to a peptide derived from a tumor antigen, a novel antigen, or an autoantigen.

[0033] In some embodiments, the receptor expresses MHC class I molecules. In some embodiments, the receptor expresses MHC class II molecules. In some embodiments, the MHC molecules stimulate an immune response to an antigen. In some embodiments, the antigen is associated with at least one disease, disorder, or condition selected from the group consisting of cancer, autoimmunity, and infection.

[0034] Various embodiments of the present invention described herein provide a method for in vitro production of platelets, the method comprising: transfecting a plurality of induced pluripotent stem cell (iPSC) precursors in an expression system, wherein the expression system is induced by a drug not present in the iPSCs; establishing a megakaryocyte precursor line by contacting the expression system with the drug to proliferate megakaryocytes; manipulating the megakaryocytes to have at least one mutation, not limited to, insertion of a nucleic acid sequence encoding the aforementioned chimeric platelet receptor; insertion of a nucleic acid sequence encoding a toxin or a protein or peptide or RNA, e.g., mRNA, e.g., cargo, e.g., a therapeutic agent or contrast agent; and deletion of a nucleic acid sequence encoding the platelet receptor; and removing the drug from the expression system to induce differentiation of the megakaryocytes into platelets.

[0035] In some embodiments, the mutation results in platelets with less immunogenicity compared to platelets from a human donor. In some embodiments, the platelets do not function similarly to platelets derived from a human donor. In some embodiments, the deletion prevents toxin release or cargo release in response to a platelet activation signal. In some embodiments, the toxin or cargo is attached to an α-granule localization signal. In some embodiments, the α-granule localization signal. In some embodiments, the method of platelet production further comprises platelets, cargo, e.g., a protein or peptide or RNA, e.g., mRNA, e.g., a therapeutic agent or contrast agent or small molecule, a toxin, and a small molecule drug, under conditions that promote absorption by platelets. In some embodiments, the expression system further comprises a platelet-specific promoter.

[0036] Various embodiments of the present invention described herein provide methods for in vivo gene editing or gene therapy in a subject, the method comprising administering engineered platelets to a subject comprising a chimeric platelet receptor described herein that is specific to the tissue to be edited, wherein the engineered platelets are coated with an adenovirus loaded with a genomic engineering mechanism, and releasing the genomic engineering mechanism in the tissue. In some embodiments, the genomic mechanism is a CRISPR / Cas gene editing system.

[0037] The various embodiments of the present invention described herein provide the use of the aforementioned therapeutic delivery system, in which the chimeric receptor is specific to the antigen associated with the disease, disorder, or condition in the treatment of the disease, disorder, or condition of interest. In some embodiments of the use described herein, the disease, disorder, or condition is selected from the group consisting of cancer, autoimmune diseases, and infections.

[0038] In some embodiments of the use described herein, cancer is acute granulocytic leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, adenocarcinoma, adenosarcoma, adrenal carcinoma, adrenocortical carcinoma, anal carcinoma, anaplastic astrocytoma, angiosarcoma, appendiceal carcinoma, astrocytoma, basal cell carcinoma, B-cell lymphoma), bile duct cancer, bladder cancer, bone cancer, intestinal cancer, brain cancer, brainstem glioma, brain tumor, breast cancer, carcinoid tumor, cervical cancer, bile duct Chondrosarcoma, chronic lymphocytic leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuse astrocytoma, ductal carcinoma in situ, endometrial cancer, ependymoma, epithelioid sarcoma, esophageal cancer, Ewing's sarcoma, extrahepatic cholangiocarcinoma, eye cancer, fallopian tube cancer, fibrosarcoma, gallbladder cancer, stomach cancer, gastrointestinal cancer, gastrointestinal carcinoid cancer, gastrointestinal stromal tumor, systemic, germ cell tumor, glioblastoma multiforme Cell tumors, gliomas, pilocytic cell leukemia, head and neck cancers, hemangioendotheliomas, Hodgkin lymphomas, Hodgkin's disease, Hodgkin lymphomas, hypopharyngeal cancers, invasive ductal carcinomas, invasive lobular carcinomas, inflammatory breast cancers, colon cancers, intrahepatic cholangiocarcinomas, invasive / invasive breast cancers, islet cell carcinomas, jaw cancers, Kaposi's sarcoma, kidney cancers, laryngeal cancers, leiomyosarcomas, leukemia, lip cancers, liposarcomas, liver cancers, lobular carcinomas in situ, low-grade astrocytosis Cancer, lung cancer, lymph node cancer, lymphoma, male breast cancer, medullary carcinoma, medulloblastoma, melanoma, meningioma, Merkel cell carcinoma, mesenchymal chondrosarcoma, mesenchymal mesothelioma, metastatic breast cancer, metastatic melanoma, metastatic squamous cell carcinoma of the neck, mixed glioma, oral cancer, mucinous carcinoma, mucosal melanoma, multiple myeloma, nasal cavity cancer, nasopharyngeal cancer, neck cancer, neuroblastoma, neuroendocrine tumor, non-Hodgkin lymphoma, non-Hodgkin's lymphoma, non-small cell lung cancer, ocular cancer, ocular melanoma, oligodendroglioma, oral cancer, oral cavity cancerCancer, oropharyngeal cancer, osteogenic sarcoma, osteosarcoma, ovarian cancer, ovarian epithelial carcinoma, ovarian germ cell tumor, primary ovarian peritoneal cancer, ovarian cord-stromal tumor, Paget's disease, pancreatic cancer, papillary carcinoma, paranasal sinus cancer, parathyroid cancer, pelvic cancer, penile cancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pilocytic cell astrocytoma, pineal gland tumor, pineoblastoma, pituitary cancer, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell carcinoma, renal pelvis cancer, rhabdomyosarcoma, salivary gland cancer, sarcoma, bone sarcoma, soft tissue sarcoma, uterine sinus cancer, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, spinal cord cancer, spinal column cancer, spinal cord cancer (Spinal cord This may include, but is not limited to, cancer, spinal tumors, squamous cell carcinoma, gastric cancer, synovial sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma / thymic carcinoma, thyroid cancer, tongue cancer, tonsil cancer, metastatic cell carcinoma, metastatic cell carcinoma, triple-negative breast cancer, fallopian tube cancer (tubal cancer), tubular carcinoma, ureteral cancer, urethral cancer, urethral cancer, uterine adenocarcinoma, uterine cancer, uterine sarcoma, vaginal cancer, and vulvar cancer.

[0039] In some embodiments of the use described herein, the disease, disorder, or condition is autoimmune disseminated encephalomyelitis, autoimmune inner ear disease, Batten disease / neuroceroid lipofuscinosis, chronic inflammatory demyelinating polyneuropathy, ophthalmosinophilic encephalitis, anti-basal ganglia, Guillain-Barré syndrome, Hashimoto's encephalopathy, anti-TPO, Isaac syndrome / acquired neurogenic myotonia, Miller-Fischer syndrome, Morvan syndrome, multiple sclerosis, gravitational myasthenia gravis, narcolepsy-pandus, Rasmussen encephalitis, generalized rigidity syndrome, Vogt-Ko Yanagi-Harada disease, Addison's disease, autoimmune hypoparathyroidism, autoimmune hypophysitis, autoimmune oophoritis, autoimmune orchitis, polyglandular autoimmune syndrome I (APECED), polyglandular autoimmune syndrome II, polyglandular autoimmune syndrome III, diabetes mellitus, type 1, Graves' disease, Hashimoto's autoimmune thyroiditis, immunodysregulation, polyglandular endocrine disorders, intestinal diseases, X-linked autoimmune hepatitis, type 1 autoimmune hepatitis, type 2 autoimmune hepatitis, autoimmune pancreatitis, abdominal diseases, Crohn's disease, pernicious anemia / atrophic gastritis, primary biliary Cirrhosis, primary sclerosing cholangitis, ulcerative colitis, acquired hemophilia A, antiphospholipid syndrome, autoimmune hemolytic anemia, autoimmune lymphoproliferative syndrome, autoimmune neutropenia, Evans syndrome, Felty syndrome, immune thrombocytopenic purpura, polymyositis / dermatomyositis, relapsing polychondritis, rheumatoid arthritis, Still's disease, alopecia areata, bullous pemphigoid, scarring pemphigoid, herpetiform dermatitis, discoid lupus erythematosus, acquired epidermolysis bullosa, linear scleroderma, pemphigus foliaceus, pemphigus vulgaris, vitiligo, Behce's disease Autoimmune diseases such as, but not limited to, To's disease, Churg-Strauss syndrome, Cogan syndrome, Crest syndrome, antifibrillarin, essential mixed cryoglobulinemia, mixed connective tissue disease, POEMS syndrome, scleroderma, Sjögren's syndrome, systemic lupus erythematosus, persistent erythema elevata, Kawasaki disease, microscopic polyangiitis, polyarteritis nodosa, rheumatic fever, Takayasu's arteritis, temporal arteritis, Wegener's granulomatosis, HLA-B27-associated acute anterior uveitis, sympathetic ophthalmitis, and Goodpasture disease.

[0040] Various embodiments of the present invention as described herein provide a therapeutic delivery system comprising: (a) engineered platelets that present a chimeric platelet receptor, the engineered platelets being produced by genetically modifying precursor megakaryocytes to be nonthrombotic and non-immunogenic; and (b) at least one therapeutic agent selected from the group consisting of cargoes, toxins, proteins, small molecule drugs, and nucleic acids packaged in vesicles within platelets as defined herein, i) the therapeutic agent being a nucleic acid or a protein, and loading occurring via expression in precursor megakaryocytes, or ii) the therapeutic agent being loaded by incubating engineered platelets with the therapeutic agent.

[0041] I. Introduction Genetically modified platelets, generated in vitro, can be targeted to release therapeutic agents (e.g., small molecule drugs, antibodies, or nucleic acids encoding proteins) in response to specific proteins expressed in target tissues or cells (e.g., cancer). These modified platelets would become targeted drug delivery devices. Unlike transplanting other tissues, platelets need to limit matching with the recipient's immune system, so modified platelets could function as "off-the-shelf" devices without needing to be matched to a specific patient (this is a major problem in current cell therapy).

[0042] The engineered platelets described herein can be generated in vitro from megakaryocytes. Because megakaryocytes are maintained in in vitro culture, they can be extensively edited at the genomic level (e.g., by CRISPR / Cas9) without fear of oncogenic transformation of the patient, which is not possible with other competing cell therapy products. Engineered platelets have an in vivo lifespan of 7–10 days and have little to no possibility of continuous reproduction, and therefore have little to no possibility of forming tumors themselves.

[0043] Platelets can be frozen and stored for long periods, resulting in a longer shelf life. Furthermore, using currently available technologies, engineered platelets can be manufactured, stored, transported, and administered to patients without issue because they are non-immunogenic.

[0044] Manipulated platelets, through in vitro genome editing of megakaryocytes, can have all thrombus-forming potential removed, mitigating potential thrombosis safety concerns. Also known as the SYNLET® therapeutic delivery system, manipulated platelets function as blank templates, effectively act as scaffolds, and possess the ability to store cargo within vesicles. Genetic alterations of megakaryocytes allow manipulated platelets to respond to specific antigens or signals. Because platelets contain a variety of signaling pathways, the manipulated input can be positive or negative, allowing the manipulated platelets to integrate and calculate various stimuli before deciding on activation. Interestingly, platelets also have the ability to transfer RNA to nearby cells, offering the possibility of genetically engineering target cells. For example, the CRISPR / Cas9 system can be delivered to target cells by manipulated platelets. Furthermore, because signaling pathways within platelets can trigger de novo translation of proteins, these can be utilized to locally deliver potentially toxic proteins to target sites.

[0045] In contrast to chimeric antigen receptor T (CAR-T) cells, the present invention provides engineered platelets, which are a universal product that does not require patient matching before administration. Furthermore, in vitro platelet production from the precursor described herein eliminates the need for continuous virus production and cell editing. Due to the short lifespan of the engineered platelets described herein, safety concerns are limited compared to current gene editing therapies. For example, since platelets are enucleated, gene editing and genomic stability are less of a concern than with CAR-T cells, and therefore the complexity of platelet therapy is not limited by editing efficiency or culture time limitations. In addition, due to their smaller size, engineered platelets may have better access to solid tumors than CAR-T cells.

[0046] Enucleated red blood cells, such as those commercially available from Rubius Therapeutics, Inc., have also been considered in the art for the delivery of therapeutic agents. In contrast to red blood cells, the engineered platelets described herein are metabolically highly active and contain a re-engineerable signaling system. In fact, engineered platelets allow for more targeted use compared to red blood cells. Vesicular degranulation of platelets also allows for the "hiding" of proteins until the target of interest is involved, which is not possible in enucleated red blood cells because biotherapeutic proteins are generally expressed on the cell surface. Thus, in one embodiment, binding of the targeting domain of a chimeric platelet receptor to a target or antigen results in degranulation of platelets containing the chimeric platelet receptor.

[0047] The manipulated platelets described herein are smaller than red blood cells and are likely to result in better biodistribution.

[0048] II. Gene Nomenclature In this specification, gene symbols are used together with ENSEMBL gene IDs to refer to human-derived genes. Unless otherwise specified, the gene names and ENSEMBL gene (ENSG) IDs corresponding to each gene symbol are shown in Table 1. The unique identifier for each ENSEMBL entry has been modified by removing the first five leading zeros (0) of the identifier after the ENSG label. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0049] In this specification, symbols and names are used together with ENSEMBL protein IDs to refer to human-derived proteins. Unless otherwise noted, Table 2 shows the protein name (as used to refer to a protein in this specification), the corresponding symbol, and the ENSEMBL protein (ENSP) ID. The unique identifier for each ENSEMBL entry has been modified by removing the first five leading zeros (0) of the identifier after the ENSP label. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6]

[0050] CD3, or CD3, is also known as differentiated cluster 2 (multiple subunits). FCER2 or CD23 is also known as the IgE receptor. NT5E is also known as 5'-nucleotidase. F9, F10 are also known as activated F9, F10. ACVRL1 is also known as activin receptor-like kinase 1. AFP is also known as alpha-fetoprotein. ANGPTL3 is also known as angiopoietin 3. BSG or CD147 is also known as basidine. APP or N / a is also known as beta-amyloid. CALCA is also known as calcitonin gene-related peptide. CA9 is also known as carbonic anhydrase 9 (CA-IX). MYH7 is also known as cardiac myosin. MET is also known as c-Met. F3 is also known as coagulation factor III. CLEC6A is also known as dendritic cell-associated lectin 2. EGFR or EGFR is also known as elongation growth factor receptor. ENG is also known as endoglin. EPHA3 is also known as ephrin receptor A3. It is also known as FGB or fibrin II, beta chain. FN1 is also known as fibronectin extradomain B. FOLH1 is also known as folate hydrolase. FOLR2 is also known as folate receptor 2. FOLR1 is also known as folate receptor alpha. FZD1 is also known as Frizzled receptor. B4GALNT1 is also known as GD2 ganglioside. ST8SIA1 is also known as GD3 ganglioside. MMP9 is also known as gelatinase B. TYRP1 or TYRP1 is also known as glycoprotein 75. GPC3 is also known as glypican 3. CSF2RA is also known as GMCSF receptor α chain. IGF1R or CD221 is also known as IGF-1 receptor. IL31RA is also known as IL31RA. ITGA2B or CD41 is also known as integrin α-IIb. ITGA5 is also known as integrin α5.ITGB3 is also known as integrin αIIbβ3. ITGB7 is also known as integrin β7. IFNg is also known as interferon gamma. IFNAR1 and IFNAR2 are also known as interferon α / β receptors. CXCL10 is also known as interferon gamma-inducing protein. IL12A or IL-12 is also known as interleukin 12. IL13 or IL-13 is also known as interleukin 13. IL17A or IL17A is also known as interleukin 17 alpha. IL17F or IL17F is also known as interleukin 17F. IL2 or IL2 is also known as interleukin 2. IL22 or IL-22 is also known as interleukin 22. IL23A or IL23 is also known as interleukin 23. IL6 or IL6 is also known as interleukin 6. SELL or CD62L is also known as L-selectin. MSLN is also known as mesoserine. MUC1 is also known as mucin CanAg. MADCAM1 is also known as mucosal adrenergic cell adhesion molecule. MAG is also known as myelin-associated glycoprotein. NECTIN4 is also known as nectin-4. CASP2 is also known as neuronal apoptosis-regulating proteinase 2. PTDSS1 is also known as phosphatidylserine. PDGFRB is also known as platelet-derived growth factor receptor beta. RHD, RHCE are also known as rhesus monkey factors. RSPO3 is also known as root plate-specific spongein 3. SELP is also known as selectin P. SAA1 or SAA2 is also known as serum amyloid A protein. APCS is also known as serum amyloid P component. S1PR1 is also known as sphingosine-1-phosphate. MAPT is also known as tau protein. TNC is also known as tenascin C. TNFRSF12A is also known as the TWEAK receptor. VIM is also known as vimentin.VWF is also known as von Willebrand factor. IL2RA or CD25 is also known as the α chain of the IL-2 receptor.

[0051] III. Compositions of the present invention Various embodiments of the present invention described herein provide megakaryocyte precursors engineered to encode chimeric platelet receptors (CPRs). These receptors can bind to specific antigens or targets present in tumors or at specific locations within the body; for example, the antigen to which the CPRs bind may be an endogenous antigen. In some embodiments, the target is not collagen.

[0052] Alternatively, platelet receptors can be removed to prevent the release of cargo or toxins in response to normal platelet activation signals. The multiple edits required to produce these precursors are possible to ensure that the progenitor cells never enter the body of a patient with ongoing culture or genomic instability concerns. Only enucleated platelets are injected into the patient.

[0053] A. Manipulation of megakaryocytes In some embodiments, the manipulated platelets described herein are derived from genetically modified megakaryocytes. The genomes of these megakaryocytes may contain knockouts of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or at least 10 genes encoding endogenous receptors, mediator proteins, and / or signaling proteins. In some cases, it is clear that it is not necessary to knock out or delete an entire gene. For example, GP1b knockout may result in abnormal platelets, but only the extracellular domain of the receptor may be deleted (deleting its ability to function) while the intracellular domain is retained, resulting in typical platelets lacking the ability to bind to von Willebrand factor, the GP1b target. Thus, in some embodiments, the disruption, deletion, or knockout described herein is a complete disruption, deletion, or knockout of an entire gene. In other embodiments, disruption, deletion, and knockout are disruptive deletions and functional knockouts, i.e., disruption of the function of a protein, and in some embodiments, deletion is a deletion of the extracellular domain of a protein.

[0054] Table 3 shows examples of genes that may be deleted from the megakaryocyte genome. [Table 3]

[0055] Those skilled in the art will understand that there are several pathways that should be disrupted in order to enable the production of platelets with a reduced likelihood of thrombus formation. In some embodiments, one or more of the following three pathways are disrupted: recognition of primary stimuli of thrombus formation, recognition of secondary stimuli of thrombus formation, and release of secondary mediators of thrombus formation.

[0056] The recognition of primary thrombus stimuli involves platelets that recognize factors related to exposed tissue that become exposed during a wound, such as recognizing the subendothelium. Under normal circumstances, platelets are not exposed to the subendothelium. Exposure to the subendothelium triggers platelets to recognize GPIb / V / IX and GPVI(GP6), ITGA2B, CLEC2, and integrin αIIb Receptors on the platelet surface, such as β3, α2β1, α5β1, and α6β1, can recognize ligands such as collagen, von Willebrand factor, fibronectin, and thrombospondin. Therefore, in some embodiments, genes encoding proteins involved in the recognition of primary stimuli for thrombus formation include GPIb / V / IX and GPVI(GP6), ITGA2B, CLEC2, and integrin α IIb This includes β3, α2β1, α5β1, and α6β1.

[0057] When platelets come into contact with exposed endothelium, for example through the above interactions, they release secondary messengers such as ADP, thrombin, and TxA2, which are detected by other platelets and cause platelet aggregation at the wound site. In some embodiments, it is preferable that the platelets' ability to recognize secondary messengers is disrupted. It is undesirable for the platelets of the present invention to target the wound site, for example, rather than the intended target. Therefore, in preferred embodiments, the platelets' ability to recognize secondary messengers is disrupted. Receptors involved in this function include Par1, Par4, P2Y12, GPIb / V / IX, thromboxane receptor (TBXA2R), P2Y1, P2X1, and integrin α IIb It contains β3.

[0058] As described above, when platelets recognize exposed tissue, they release secondary messengers to recruit other platelets to that site. When the platelets of the present invention bind to a target, such as a tumor antigen, it is undesirable for the platelets of the present invention to then recruit other platelets to the target site to form a thrombus, such as a thrombus at the tumor site. Therefore, in a preferred embodiment, the pathway by which activated platelets release secondary messengers is disrupted. The pathway may include proteins involved in the production and / or storage and / or release of secondary mediators. Genes involved in this pathway include Cox1, HPS, and thromboxane A synthase (TBXAS1).

[0059] Those skilled in the art will understand that a single gene may be involved in two or three of the functions described above.

[0060] In some embodiments, it is more preferable that megakaryocytes or their precursors contain disruptions or deletions (e.g., knockouts) of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or at least 10 genes encoding proteins involved in the recognition of primary stimuli of thrombosis, proteins involved in the recognition of secondary mediators of thrombosis, and / or proteins involved in the release of secondary mediators of thrombosis.

[0061] It will be apparent to those skilled in the art that the meaning of any protein involved in this process is included by the proteins involved in the recognition of the primary stimulus of a thrombus, for example, proteins directly involved in contact with or recognition of the primary stimulus of a thrombus, and also, for example, genes that lead to the expression of those proteins directly involved in contact with or recognition of the primary stimulus of a thrombus. Those skilled in the art will understand which proteins are considered to be involved in the recognition of the primary stimulus. An important feature is that the disruption of these proteins leads to a defect in the recognition of the primary stimulus of a thrombus. However, in some embodiments, the proteins involved in the recognition of the primary stimulus of a thrombus include only proteins that directly contact the primary stimulus of a thrombus.

[0062] Proteins involved in the recognition of secondary mediators of thrombosis include proteins directly involved in contact with or recognition of secondary mediators of thrombosis, as well as proteins indirectly involved in these processes, such as proteins involved in the production of proteins directly involved in contact with or recognition of secondary mediators of thrombosis. Those skilled in the art will understand what it means for proteins involved in the recognition of secondary mediators of thrombosis. A key characteristic of these proteins is that their disruption results in a defect in the recognition of secondary mediators of thrombosis. However, in some embodiments, proteins involved in the recognition of secondary mediators of thrombosis include only proteins that directly contact secondary mediators of thrombosis.

[0063] The proteins involved in the release of secondary mediators in thrombus formation include those involved in the production and / or storage and / or release of secondary mediators. A key characteristic of these proteins is that their disruption results in defects in the final release of secondary mediators. These defects may lie in the production, storage, and / or actual release processes of secondary mediators.

[0064] In some embodiments, megakaryocytes or their precursors are at least: One gene encoding a protein involved in recognizing the primary stimulus for thrombus formation, One gene encoding a protein involved in the recognition of secondary mediators of thrombus formation, and This involves disruption or deletion of one gene that encodes a protein involved in the release of secondary mediators of thrombus formation. In some embodiments, megakaryocytes or their precursors are at least: Two genes encoding proteins involved in the recognition of the primary stimulus for thrombus formation, Two genes encoding proteins involved in the recognition of secondary mediators of thrombus formation, and This involves disruption or deletion of two genes that encode proteins involved in the release of secondary mediators of thrombus formation. In some embodiments, megakaryocytes or their precursors are at least: Three genes encoding proteins involved in the recognition of the primary stimulus for thrombus formation, Three genes encoding proteins involved in the recognition of secondary mediators of thrombus formation, and This involves disruption or deletion of three genes that encode proteins involved in the release of secondary mediators of thrombus formation. Genes thought to encode proteins involved in the recognition of primary stimuli for thrombus formation include GPIb / V / IX and GPVI (GP6), ITGA2B, CLEC2, and integrin s α IIbβ3, α2β1, α5β1, and α6β1, or optionally GPVI and ITGA2B. Genes thought to encode proteins involved in recognizing secondary stimuli for thrombosis include Par1, Par4, P2Y12, GPIb / V / IX, thromboxane receptor (TBXA2R), P2Y1, P2X1, and integrin α IIb β3, or optionally Par1, Par4, and P2Y12. Genes thought to encode proteins involved in releasing secondary mediators of thrombosis include Cox1, HPS, and thromboxane A synthase (TBXAS1), or optionally Cox1 and HPS.

[0065] In some embodiments: At least one, two, or three genes encoding proteins involved in recognizing primary stimuli for thrombosis are selected from the group consisting of GPIb / V / IX and GPVI (GP6), ITGA2B, CLEC2, integrin s α IIb β3, α2β1, α5β1, and α6β1, or selected from the group consisting of GPVI and ITGA2B, At least one, two, or three encoding proteins involved in recognizing secondary mediators of thrombosis are selected from the group consisting of Par1, Par4, P2Y12, GPIb / V / IX, thromboxane receptor (TBXA2R), P2Y1, P2X1, and integrin α IIb β3, or selected from the group consisting of Par1, Par4, and P2Y12, and / or At least one, two, or three genes encoding proteins involved in releasing secondary mediators of thrombosis are selected from the group consisting of Cox1, HPS, and thromboxane A synthase (TBXAS1), or selected from the group consisting of Cox1 and HPS.

[0066] In a preferred embodiment, the genetically modified megakaryocytes or their precursors are the following genes: GPVI, ITGA2B, Par1, Par4, P2Y12, Cox1, and HPS each have defects or missing data.

[0067] For example, genetically modified megakaryocytes or their precursors may include knockouts of GPVI, ITGA2B, Par1, Par4, P2Y12, Cox1, and HPS.

[0068] In some embodiments, the expression of the genes in Table 3 may be altered or “knocked out” using the CRISPR / Cas system, zinc finger nucleases, transcriptional activator-like effector nucleases (TALENs), RNA interference constructs (RNAi) (e.g., small interfering RNA (siRNA) or microRNA (miRNA), or short hairpin RNA (shRNA)).

[0069] The effects of megakaryocyte gene knockouts on the resulting engineered platelets can vary. For example, the RAB27a (RAS oncogene) and HPS (haptoglobin) genes function in dense granule loading and formation, respectively. Knockout or deletion of Rab27a may result in engineered platelets that lack dense granule mediators but otherwise exhibit normal platelet biology. Knockout or deletion of the HPS gene may result in engineered platelets that do not contain dense granules. Knockout or deletion of AIIbB3 or GP1b-IX-V may reduce the interaction between activated platelets and von Willebrand factor (vWF), potentially preventing platelets from aggregating with each other. Furthermore, AIIbB3 is also involved in inverse signaling to enhance integrin affinity for fibrinogen (see Durrant, Blood. 2017 Oct 5;130(14):1607-1619). Knockout or deletion of IP (PGI2R or prostaglandin I2 receptor) may result in negative regulation of prostaglandins. Knockout or deletion of TP (TxA2R or thromboxane A2 receptor) may result in additional platelet recruitment upon activation to stimulate coagulation.

[0070] GPVI (ITAM receptor) has also been observed to be stimulated in G protein α-q (Galphaq) knockout mice. Conversely, ITAM agonists such as collagen induce the release of ADP and G protein-coupled receptors (GPCR agonists) such as thromboxane A2 receptor (TXA2), indirectly activating phospholipase C (PLC) via the Gq pathway. Furthermore, Galphaq is active for the proper functioning of thrombin, ADP, 5-hydroxytryptamine (5HT), PAF, and thromboxane A (TXA).

[0071] Knockout or deletion of P-selectin, thromboxane synthase, and platelet-activating factor (PAF) may result in impaired platelet aggregation once activated. Knockout or deletion of LIM domain kinase (LIMK1) may reduce TxA2 synthesis. CXCL4 (CXC-motif chemokine ligand 4) and CXCL7 (CXC-motif chemokine ligand 7) are chemokines, and therefore, knockout or deletion of these genes may interfere with at least one signaling pathway. Tallin 1 and kindlin function in signaling, allowing integrins to enter a sensitive state.

[0072] Knockout or deletion of ANO6 / TMEM16F disrupts the platelet's ability to expose itself to phosphatidylserine upon platelet activation. Phosphatidylserine is a membrane lipid normally retained on the cytoplasmic surface of platelets. Upon platelet activation, calcium influx triggers phosphatidylserine exposure on the platelet's outer surface via ANO6 / TMEM16F, which, in combination with coagulation factors, catalyzes the production of activated thrombin. Therefore, knockout of TMEM16F prevents phosphatidylserine exposure and reduces the thrombogenicity of platelets. This is exemplified in patients with Scott syndrome, characterized by ANO6 mutations and a clinically high risk of bleeding.

[0073] B. Chimeric platelet receptor (CPR) In some embodiments, the engineered platelets described herein may include modifications to endogenous platelet receptors. Modifications include, but are not limited to, deletion or addition of the entire receptor or domain of these receptors, or combination with domains from non-endogenous receptors, resulting in differences in the behavior of engineered platelets compared to unmodified platelets. Domains from immunoreceptor tyrosine-based activation motifs (ITAMs) receptors can be used in chimeric platelet receptors to stimulate platelet activation. This inhibits the activation of platelet immunoreceptor tyrosine-based inhibitory motifs (ITIMs).

[0074] Since the ITAM domain can still activate the same downstream signaling components that the ITAM receptor endogenously finds in platelets, it will be apparent to those skilled in the art that a domain from the ITAM receptor, which is not normally expressed in platelets, still functions in the present invention.

[0075] 1. Endogenous platelet receptors In some embodiments, platelets may be redirected to degranulate by antigens rather than collagen. ITIM-containing receptors directly counteract the activation of ITIM receptors by inhibiting platelet activation. CEACAM-1, PECAM-1, and G6b-B are ITIM-containing receptors. Antibody-mediated G6b-B clustering inhibits platelet activation via GPVI and CLEC-2, as shown in Mori et al. “G6b-B inhibits constitutive and agonist-induced signaling by glycoprotein VI and CLEC-2”. JBC, 2008, which is incorporated herein in whole by reference. Adding chimeric “off” receptors may be used to improve the specificity of the synthetic platelets described herein. Manipulated chimeric immunoreceptor tyrosine-based inhibitory motif (ITIM) receptors enable the construction of logic gates.

[0076] Alternatively, the ITAM receptor mediates platelet activation and stimulates an immune response. Glycoprotein VI (GPVI) binds to collagen and is a central mediator of platelet activation. Characterized by an extracellular IgG-like domain, its internal tyrosine kinase signaling pathway is triggered by receptor clustering via the Fc receptor (FcR) gamma chain. In certain embodiments, the intracellular domain is retained, and the extracellular domain is exchanged to target antigens. For example, in some embodiments, the chimeric platelet receptor contains an intracellular domain that stimulates platelets but does not contain a corresponding extracellular domain. For example, in some embodiments, the extracellular targeting domain of the receptor is heterogeneous to the intracellular domain of the receptor. A heterogeneous extracellular targeting domain means that the extracellular domain is not the usual extracellular domain associated with the intracellular domain. For example, in embodiments where the intracellular domain contains the extracellular domain of glycoprotein VI (GPVI), the extracellular domain is not the extracellular domain of glycoprotein VI (GPVI), and the domains are heterogeneous to each other.

[0077] Alternatively, C-type lectin-like receptor 2 (CLEC-2) or the Fc fragment of IgG receptor IIa (FCgR2A) can be modified in a similar manner. In other embodiments, if the intracellular domain includes the intracellular domain of C-type lectin-like receptor 2 (CLEC-2), the extracellular targeting domain is not the extracellular domain of CLEC-2, and in some embodiments, if the intracellular domain includes the Fc fragment of IgG receptor IIa (FCgR2A), the extracellular targeting domain does not include the extracellular domain of FCgR2A. It is clear that the extracellular targeting domain may be a domain specific to the target but not specific to the intracellular domain.

[0078] In that case, it would be clear that in some embodiments, CPR is not a naturally occurring protein.

[0079] CLEC-2 binds to podoplanin (associated with tumors) and, in response to snake venom rhodocytin, induces platelet activation, thereby inducing platelet aggregation through the activation of Src and Syk non-receptor tyrosine kinases in the internal tyrosine kinase signaling pathway, triggered by receptor clustering via the signaling protein lymphocyte cytoplasmic protein 2 or 76 kDa SH2 domain-containing leukocyte protein (SLP-76) and 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterase gamma-2 (PLCγ2) (see Fu et al. Blood, 127(13):1629-30, 2016, the entire information of which is incorporated herein by reference).

[0080] FCgr2A binds to the Fc region of antibodies and triggers platelet activation in response to opsonizing bacteria via the signaling protein Syk non-receptor tyrosine kinase. The internal tyrosine kinase signaling pathway is triggered by receptor clustering.

[0081] In some embodiments, additional ITAM receptors can be inserted into the megakaryocyte genome to enhance T cell signaling and stimulate an immune response. T cell receptors (TCRs) recognize antigens bound to the major histocompatibility complex (MHC) (see James et al. Sci.Signal.11,eaan1088 (2018), the entire structure of which is incorporated herein by reference). ITAM on TCRs translates the binding and recognition actions into intracellular signals (ibid.). Insertion of additional ITAM into chimeric TCRs was observed to scale proportionally to the number of ITAM receptors, and reduction or knockout of the number of ITAM receptors was observed to inhibit T cell development by impairing the involvement of thymocyte lineages (ibid.).

[0082] In some embodiments, the CPR may comprise one or more domains or portions thereof from one or more immunoreceptor tyrosine-based activation motif (ITAM) receptors. Non-limiting examples of ITAM receptors include glycoprotein VI platelet (GPVIA), high affinity immunoglobulin epsilon receptor subunit gamma (FCERG), type C lectin domain family 1 (CLEC1), and the Fc fragment of IgG receptor II (FCGR2).

[0083] In one embodiment, the domains of the ITAM receptors FCERG (SEQ ID NO: 1), CLEC1 (SEQ ID NO: 6), FCGR2 (SEQ ID NO: 10), and / or GPVIA (SEQ ID NO: 15), as shown in Table 4, can be combined for expression in megakaryocytes to produce CPR in manipulated platelets.

[0084] In one embodiment, the domain of the ITAM receptor can be combined with the domain of the T cell receptor to form a chimeric ITAM receptor, also known as a chimeric platelet receptor. These chimeric receptors can be combined for expression in megakaryocytes to induce CPR in engineered platelets. Table 4 shows non-limiting examples of chimeric ITAM receptors FCERG (SEQ ID NO: 20), CLEC1 (SEQ ID NO: 21), FCGR2 (SEQ ID NO: 22), and GPVIA (SEQ ID NO: 23). [Table 4]

[0085] SEQ ID NO: 1 is an embodiment of the complete FCERG receptor. SEQ ID NO: 2 is an embodiment of the signal peptide of FCERG. SEQ ID NO: 3 is an embodiment of the extracellular domain of FCERG. SEQ ID NO: 4 is an embodiment of the transmembrane domain of FCERG. SEQ ID NO: 5 is an embodiment of the cytoplasmic domain of FCERG. SEQ ID NO: 6 is an embodiment of the ITAM receptor of CLEC1. SEQ ID NO: 7 is an embodiment of the cytoplasmic domain of CLEC1. SEQ ID NO: 8 is an embodiment of the transmembrane domain of CLEC1. SEQ ID NO: 9 is an embodiment of the extracellular domain of CLEC1. SEQ ID NO: 10 is an embodiment of the ITAM receptor of FCGR2. SEQ ID NO: 11 is an embodiment of the signal peptide of FCGR2. SEQ ID NO: 12 is an embodiment of the extracellular domain of FCGR2. SEQ ID NO: 13 is an embodiment of the transmembrane domain of FCGR2. SEQ ID NO: 14 is an embodiment of the cytoplasmic domain of FCGR2. SEQ ID NO: 15 is an embodiment of the ITAM receptor of GPVIA. SEQ ID NO: 16 is an embodiment of the signal peptide of GPVIA. Sequence ID 17 is an embodiment of the extracellular domain of GPVIA. Sequence ID 18 is an embodiment of the transmembrane domain of GPVIA. Sequence ID 19 is an embodiment of the cytoplasmic domain of GPVIA.

[0086] Sequence ID 20 is an embodiment of a chimeric ITAM receptor based on FCERG. Sequence ID 21 is an embodiment of a chimeric ITAM receptor based on CLEC1. Sequence ID 22 is an embodiment of a chimeric ITAM receptor based on FCGR2. Sequence ID 23 is an embodiment of a chimeric ITAM receptor based on GPVIA.

[0087] In some embodiments, the CPR may comprise one or more domains or portions thereof from one or more immunoreceptor tyrosine-based inhibitory motif (ITIM) receptors. Non-limiting examples of ITIM receptors include platelet and endothelial cell adhesion molecule 1 (PECAM1), trigger receptor expressed in myeloid cells 1 (TLT1), leukocyte immunoglobulin-like receptor B2 (LILRB2), carcinoembryonic antigen-associated cell adhesion molecule 1 (CEACAM1), and megakaryocyte and platelet inhibitory receptor G6b (G6b-B).

[0088] In one embodiment, the domains of the ITIM receptors LILRB2 (SEQ ID NO: 34), PECAM1 (SEQ ID NO: 38), TLT1 (SEQ ID NO: 43), and CEACAM1 (SEQ ID NO: 24), shown in Table 5, are combined for expression in megakaryocytes to result in CPR of engineered platelets.

[0089] In one embodiment, the ITIM receptor domain can be combined with the T cell receptor domain to form a chimeric ITIM receptor, also known as a chimeric platelet receptor. These chimeric receptors can be combined for expression in megakaryocytes to induce CPR in engineered platelets. [Table 5]

[0090] Sequence ID 24 is an embodiment of the ITIM receptor of CEACAM1. Sequence ID 25 is an embodiment of the signal peptide of CEACAM1. Sequence ID 26 is an embodiment of the extracellular domain of CEACAM1. Sequence ID 27 is an embodiment of the transmembrane domain of CEACAM1. Sequence ID 28 is an embodiment of the cytoplasmic domain of CEACAM1. Sequence ID 29 is an embodiment of the ITIM receptor of G6b-B. Sequence ID 30 is an embodiment of the signal peptide of G6b-B. Sequence ID 31 is an embodiment of the extracellular domain of G6b-B. Sequence ID 32 is an embodiment of the transmembrane domain of G6b-B. Sequence ID 33 is an embodiment of the cytoplasmic domain of G6b-B. Sequence ID 34 is an embodiment of the ITIM receptor of LILRB2. Sequence ID 35 is an embodiment of the signal peptide of LILRB2. Sequence ID 36 is an embodiment of the additional domain of LILRB2. Sequence ID 37 is an embodiment of the transmembrane domain of LILRB2. Sequence ID 38 is an embodiment of the ITIM receptor of PECAM1. Sequence ID 39 is an embodiment of the signal peptide of PECAM1. Sequence ID 40 is an embodiment of the extracellular domain of PECAM1. Sequence ID 41 is an embodiment of the transmembrane domain of PECAM1. Sequence ID 42 is an embodiment of the cytoplasmic domain of PECAM1. Sequence ID 43 is an embodiment of the ITIM receptor of TLT1. Sequence ID 44 is an embodiment of the signal peptide of TLT1. Sequence ID 45 is an embodiment of the extracellular domain of TLT1. Sequence ID 46 is an embodiment of the transmembrane domain of TLT1. Sequence ID 47 is an embodiment of the cytoplasmic domain of TLT1.

[0091] a.domain CPR may include any combination of a signal peptide, an extracellular domain, a transmembrane domain, a cytoplasmic domain, or a linker or targeting domain.

[0092] In one embodiment, the CPR may include a signal peptide selected from Table 6. [Table 6]

[0093] In one embodiment, the CPR comprises at least one FCERG signal peptide. In a non-limiting example, the FCERG signal peptide is Sequence ID No. 2.

[0094] In one embodiment, the CPR comprises at least one FCGR2 signal peptide. As a non-limiting example, the FCGR2 signal peptide is SEQ ID NO: 11.

[0095] In one embodiment, the CPR comprises at least one GPVIA signal peptide. As a non-limiting example, the GPVIA signal peptide is SEQ ID NO: 16.

[0096] In one embodiment, the CPR comprises at least one CEACAM1 signal peptide. As a non-limiting example, the CEACAM1 signal peptide is Sequence ID No. 25.

[0097] In one embodiment, CPR comprises at least one G6b-B signal peptide. As a non-limiting example, the G6b-B signal peptide is SEQ ID NO: 30.

[0098] In one embodiment, the CPR comprises at least one LILRB2 signal peptide. As a non-limiting example, the LILRB2 signal peptide is SEQ ID NO: 35.

[0099] In one embodiment, the CPR comprises at least one PECAM1 signal peptide. As a non-limiting example, the PECAM1 signal peptide is SEQ ID NO: 39.

[0100] In one embodiment, the CPR comprises at least one TLT1 signal peptide. As a non-limiting example, the TLT1 signal peptide is SEQ ID NO: 44.

[0101] CPR may comprise a portion of the signal peptides in Table 6 or signal peptides known in the art. This portion may consist of 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 nucleotides of any of the sequences in Table 6, including but not limited to SEQ ID NOs: 2, 11, 16, 25, 30, 35, 39, and 44. This portion may consist of 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides of any of the sequences in Table 7, including but not limited to SEQ ID NOs: 2, 11, 16, 25, 30, 35, 39, and 44.

[0102] CPR may include an extracellular (EC) domain or a portion thereof, selected from Table 7. [Table 7]

[0103] In one embodiment, the CPR includes at least one FCERG EC domain. In a non-limiting example, the FCERG EC domain is Sequence ID No. 3.

[0104] In one embodiment, the CPR includes at least one CLEC1 EC domain. In a non-limiting example, the CLEC1 EC domain is sequence number 9.

[0105] In one embodiment, the CPR includes at least one CLEC1 EC domain. In a non-limiting example, the CLEC1 EC domain is sequence number 53.

[0106] In one embodiment, the CPR includes at least one FCGR2 EC domain. In a non-limiting example, the FCGR2 EC domain is Sequence ID No. 12.

[0107] In one embodiment, the CPR includes at least one FCGR2 EC domain. In a non-limiting example, the FCGR2 EC domain is Sequence ID No. 54.

[0108] In one embodiment, the CPR includes at least one GPVIA EC domain. In a non-limiting example, the GPVIA EC domain is Sequence ID No. 17.

[0109] In one embodiment, the CPR includes at least one GPVIA EC domain. In a non-limiting example, the GPVIA EC domain is Sequence ID No. 55.

[0110] In one embodiment, the CPR includes at least one CEACAM1 EC domain. In a non-limiting example, the CEACAM1 EC domain is Sequence ID No. 26.

[0111] In one embodiment, the CPR includes at least one G6b-B EC domain. As a non-limiting example, the G6b-B EC domain is Sequence ID No. 31.

[0112] In one embodiment, the CPR includes at least one LILRB2 EC domain. In a non-limiting example, the LILRB2 EC domain is Sequence ID No. 36.

[0113] In one embodiment, the CPR includes at least one PECAM1 EC domain. In a non-limiting example, the PECAM1 EC domain is Sequence ID No. 40.

[0114] In one embodiment, the CPR includes at least one TLT1 EC domain. In a non-limiting example, the TLT1 EC domain is Sequence ID No. 45.

[0115] The CPR may include a portion of the EC domains in Table 7, or an EC domain known in the art. This portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 nucleotides of any of the sequences in Table 7, including but not limited to sequence numbers 3, 9, 12, 17, 26, 31, 36, 40, and 45. This portion may be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides of any of the sequences in Table 7, including but not limited to sequence numbers 3, 9, 12, 17, 26, 31, 36, 40, and 45.

[0116] In one embodiment, the CPR may include a portion of the FCERG EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 3. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 3.

[0117] In one embodiment, the CPR may include a portion of the CLEC1 EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 9. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 9.

[0118] In one embodiment, the CPR may include a portion of the FCERG EC domain shown in Table 7, which is 10 to 15 nucleotides long. This portion may be nucleotides 10, 11, 12, 13, 14, or 15 of SEQ ID NO: 3.

[0119] In one embodiment, the CPR may include a portion of the CLEC1 EC domain shown in Table 7, which is 10 to 15 nucleotides long. This portion may be nucleotides 10, 11, 12, 13, 14, or 15 of SEQ ID NO: 9 or 53. As a non-limiting example, the portion of SEQ ID NO: 9 may be SEQ ID NO: 53.

[0120] In one embodiment, the CPR may include a portion of the CLEC1 EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 53. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 53.

[0121] In one embodiment, the CPR may include a portion of the FCGR2 EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 12. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 12.

[0122] In one embodiment, the CPR may include a portion of the FCGR2 EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 54. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 54.

[0123] In one embodiment, the CPR may include a portion of the GPVIA EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 17. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 17.

[0124] In one embodiment, the CPR may include a portion of the GPVIA EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 55. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 55.

[0125] In one embodiment, the CPR may include a portion of the CEACAM1 EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 26. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 26.

[0126] In one embodiment, the CPR may include a portion of the G6b-B EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 31. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 31.

[0127] In one embodiment, the CPR may include a portion of the LILRB2 EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 36. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 36.

[0128] In one embodiment, the CPR may include a portion of the PECAM1 EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 40. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 40.

[0129] In one embodiment, the CPR may include a portion of the TLT1 EC domain shown in Table 7. This portion may be nucleotides 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 of SEQ ID NO: 45. This portion may be nucleotides 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 of SEQ ID NO: 45.

[0130] CPR may include transmembrane (TM) domains selected from Table 8. [Table 8]

[0131] In one embodiment, the CPR includes at least one FCERG™ domain. In a non-limiting example, the FCERG™ domain is Sequence ID No. 4.

[0132] In one embodiment, the CPR includes at least one CLEC1™ domain. In a non-limiting example, the CLEC1™ domain is Sequence ID No. 8.

[0133] In one embodiment, the CPR includes at least one FCGR2™ domain. In a non-limiting example, the FCGR2™ domain is Sequence ID No. 13.

[0134] In one embodiment, the CPR includes at least one GPVIA™ domain. In a non-limiting example, the GPVIA™ domain is Sequence ID No. 18.

[0135] In one embodiment, the CPR includes at least one CEACAM1™ domain. In a non-limiting example, the CEACAM1™ domain is Sequence ID No. 27.

[0136] In one embodiment, the CPR includes at least one G6b-B™ domain. In a non-limiting example, the G6b-B™ domain is Sequence ID No. 32.

[0137] In one embodiment, the CPR includes at least one LILRB2™ domain. In a non-limiting example, the LILRB2™ domain is Sequence ID No. 37.

[0138] In one embodiment, the CPR includes at least one PECAM1™ domain. In a non-limiting example, the PECAM1™ domain is Sequence ID No. 41.

[0139] In one embodiment, the CPR includes at least one TLT1™ domain. In a non-limiting example, the TLT1™ domain is Sequence ID No. 46.

[0140] CPR may include cytoplasmic (cellular) domains selected from Table 9. [Table 9]

[0141] In one embodiment, the CPR includes at least one FCERG cytoplasmic domain. In a non-limiting example, the FCERG cytoplasmic domain is Sequence ID No. 5.

[0142] In one embodiment, the CPR includes at least one CLEC1 cytoplasmic domain. In a non-limiting example, the CLEC1 cytoplasmic domain is Sequence ID No. 7.

[0143] In one embodiment, the CPR comprises at least one FCGR2 cytoplasmic domain. In a non-limiting example, the FCGR2 cytoplasmic domain is Sequence ID No. 14.

[0144] In one embodiment, the CPR includes at least one GPVIA cytoplasmic domain. In a non-limiting example, the GPVIA cytoplasmic domain is Sequence ID No. 19.

[0145] In one embodiment, the CPR includes at least one CEACAM1 cytoplasmic domain. In a non-limiting example, the CEACAM1 cytoplasmic domain is Sequence ID No. 28.

[0146] In one embodiment, the CPR comprises at least one G6b-B cytoplasmic domain. As a non-limiting example, the G6b-B cytoplasmic domain is Sequence ID No. 33.

[0147] In one embodiment, the CPR includes at least one LILRB2 cytoplasmic domain. In a non-limiting example, the LILRB2 cytoplasmic domain is Sequence ID No. 52.

[0148] In one embodiment, the CPR includes at least one PECAM1 cytoplasmic domain. In a non-limiting example, the PECAM1 cytoplasmic domain is Sequence ID No. 42.

[0149] In one embodiment, the CPR includes at least one TLT1 cytoplasmic domain. In a non-limiting example, the TLT1 cytoplasmic domain is Sequence ID No. 47.

[0150] Genes encoding fusion peptides, targeting domains, or linking proteins can be added to megakaryocyte genomes, such as the light chain of variable fragment 1 having the nucleic acid sequence of SEQ ID NO: 48 (Fv1_L chain) or an improved linker from a single-chain variable fragment with reduced aggregation and enhanced proteolytic stability having the nucleic acid sequence of SEQ ID NO: 49 (Whitlow_ linker), as shown in Table 10. Alternatively, at least a portion of an antibody can be added to the megakaryocyte genome for expression in platelets, such as the kappa light chain of an anti-human B cell CD19 antibody (F1H chain_CD19FMC63) having the nucleic acid sequence of SEQ ID NO: 50 and a modified IGg4 hinge region having the nucleic acid sequence of SEQ ID NO: 51, also shown in Table 10.

[0151] In some embodiments, the CPR includes at least one domain selected from Tables 4-9 and a linker and / or targeting domain selected from Table 10. [Table 10]

[0152] In one embodiment, the CPR has a domain that has at least 95% identity with any of the sequences in Tables 4 to 10, including sequence numbers 1 to 55. In one embodiment, the CPR has a domain having 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with at least one of the sequences in Tables 4 to 10, including sequence numbers 1 to 55.

[0153] In one embodiment, the CPR has at least one signal peptide having at least 95% identity to any of the sequences in Table 6, including SEQ ID NOs: 2, 11, 16, 25, 30, 35, 39, and 44. In one embodiment, the CPR has 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, and 71% identity to at least one of the sequences in Table 6, including SEQ ID NOs: 2, 11, 16, 25, 30, 35, 39, and 44. It has at least one signal peptide domain having identity of 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0154] In one embodiment, the CPR has at least one extracellular domain having at least 95% identity to any of the sequences in Table 7, including SEQ ID NOs: 3, 9, 53, 12, 54, 17, 55, 26, 31, 36, 40, or 45. In one embodiment, the CPR has 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69% identity to at least one of the sequences in Table 7, including SEQ ID NOs: 3, 9, 53, 12, 54, 17, 55, 26, 31, 36, 40, or 45. It has at least one extracellular domain having identity of 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0155] In one embodiment, the CPR has at least one transmembrane domain having at least 95% identity to any of the sequences in Table 8, including sequence numbers 4, 8, 13, 18, 27, 32, 37, 41, or 46. In one embodiment, the CPR has 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, It has at least one transmembrane domain having identity of 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0156] In one embodiment, the CPR has at least one cytoplasmic domain having at least 95% identity to any of the sequences in Table 9, including sequence numbers 5, 7, 14, 19, 28, 33, 52, 42, or 47. In one embodiment, the CPR has 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, and 70% identity to at least one of the sequences in Table 9, including sequence numbers 5, 7, 14, 19, 28, 33, 52, 42, or 47. It has at least one cytoplasmic domain having identity of 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0157] In one embodiment, the CPR has at least one linker or targeting domain that has at least 95% identity to any of the sequences in Table 10, including sequence numbers 48, 49, 50, or 51. In one embodiment, the CPR has 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 7 It has at least one linker or targeting domain having identity of 3%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%.

[0158] In one embodiment, the CPR comprises a signal peptide, an extracellular domain, a transmembrane domain, and a cytoplasmic domain.

[0159] In one embodiment, the CPR comprises an extracellular domain, a transmembrane domain, and a cytoplasmic domain.

[0160] b. Domain swapping In one embodiment, any of the ITAM and / or ITIM receptor domains in Tables 4 and 5 may be replaced by a domain from another ITAM and / or ITIM receptor.

[0161] In one embodiment, the EC domain of the ITAM receptor shown in Table 4 may be replaced with a domain from another ITAM and / or ITIM receptor. For example, the EC domain of the FCERG ITAM receptor may be replaced with the CLEC1 EC domain or a portion thereof to generate a CPR. For example, the EC domain of the FCERG ITAM receptor may be replaced with the FCGR2 EC domain or a portion thereof to generate a CPR. For example, the EC domain of the FCERG ITAM receptor may be replaced with the GPVIA EC domain or a portion thereof to generate a CPR. For example, the EC domain of the FCERG ITAM receptor may be replaced with the LILRB2 EC domain or a portion thereof to generate a CPR. For example, the EC domain of the FCERG ITAM receptor may be replaced with the PECAM1 EC domain or a portion thereof to generate a CPR. For example, the EC domain of the FCERG ITAM receptor may be replaced with the TLT1 EC domain or a portion thereof to generate a CPR. For example, the EC domain of the FCERG ITAM receptor may be replaced with the CEACAM1 EC domain or a portion thereof to generate a CPR.

[0162] For example, the TM domain of the FCERG ITAM receptor can be replaced by the CLEC1 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCERG ITAM receptor can be replaced by the FCGR2 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCERG ITAM receptor can be replaced by the GPVIA TM domain or a part thereof to generate CPR. For example, the TM domain of the FCERG ITAM receptor can be replaced by the LILRB2 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCERG ITAM receptor can be replaced by the PECAM1 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCERG ITAM receptor can be replaced by the TLT1 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCERG ITAM receptor can be replaced by the CEACAM1 TM domain or a part thereof to generate CPR.

[0163] For example, the signal peptide of the FCERG ITAM receptor may be replaced by the CLEC1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCERG ITAM receptor may be replaced by the FCGR2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCERG ITAM receptor may be replaced by the GPVIA signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCERG ITAM receptor may be replaced by the LILRB2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCERG ITAM receptor may be replaced by the PECAM1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCERG ITAM receptor may be replaced by the TLT1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCERG ITAM receptor may be replaced by the CEACAM1 signal peptide or a part thereof to generate CPR.

[0164] For example, the cytoplasmic domain of the FCERG ITAM receptor may be replaced by the CLEC1 cytoplasmic domain or a part thereof to generate CPR. For example, the cytoplasmic domain of the FCERG ITAM receptor may be replaced by the FCGR2 cytoplasmic domain or a part thereof to generate CPR. For example, the cellular domain of the FCERG ITAM receptor may be replaced by the GPVIA cellular domain or a part thereof to generate CPR. For example, the cellular domain of the FCERG ITAM receptor may be replaced by the LILRB2 cellular domain or a part thereof to generate CPR. For example, the cytoplasmic domain of the FCERG ITAM receptor may be replaced by the PECAM1 cytoplasmic domain or a part thereof to generate CPR. For example, the cytoplasmic domain of the FCERG ITAM receptor may be replaced by the TLT1 cytoplasmic domain or a part thereof to generate CPR. For example, the cellular domain of the FCERG ITAM receptor may be replaced by the CEACAM1 cellular domain or a part thereof to generate CPR.

[0165] For example, the EC domain of the CLEC1 ITAM receptor can be replaced by the FCGR2 EC domain or a part thereof to generate CPR. For example, the EC domain of the CLEC1 ITAM receptor can be replaced by the GPVIA EC domain or a part thereof to generate CPR. For example, the EC domain of the CLEC1 ITAM receptor can be replaced by the LILRB2 EC domain or a part thereof to generate CPR. For example, the EC domain of the CLEC1 ITAM receptor can be replaced by the PECAM1 EC domain or a part thereof to generate CPR. For example, the EC domain of the CLEC1 ITAM receptor can be replaced by the TLT1 EC domain or a part thereof to generate CPR. For example, the EC domain of the CLEC1 ITAM receptor can be replaced by the CEACAM1 EC domain or a part thereof to generate CPR.

[0166] For example, the TM domain of the CLEC1 ITAM receptor can be replaced by the FCERG TM domain or a part thereof to generate CPR. For example, the TM domain of the CLEC1 ITAM receptor can be replaced by the FCGR2 TM domain or a part thereof to generate CPR. For example, the TM domain of the CLEC1 ITAM receptor can be replaced by the GPVIA TM domain or a part thereof to generate CPR. For example, the TM domain of the CLEC1 ITAM receptor can be replaced by the LILRB2 TM domain or a part thereof to generate CPR. For example, the TM domain of the CLEC1 ITAM receptor can be replaced by the PECAM1 TM domain or a part thereof to generate CPR. For example, the TM domain of the CLEC1 ITAM receptor can be replaced by the TLT1 TM domain or a part thereof to generate CPR. For example, the TM domain of the CLEC1 ITAM receptor can be replaced by the CEACAM1 TM domain or a part thereof to generate CPR.

[0167] For example, the signal peptide of the CLEC1 ITAM receptor may be replaced by the FCERG signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CLEC1 ITAM receptor may be replaced by the FCGR2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CLEC1 ITAM receptor may be replaced by the GPVIA signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CLEC1 ITAM receptor may be replaced by the LILRB2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CLEC1 ITAM receptor may be replaced by the PECAM1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CLEC1 ITAM receptor may be replaced by the TLT1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CLEC1 ITAM receptor may be replaced by the CEACAM1 signal peptide or a part thereof to generate CPR.

[0168] For example, the cellular domain of the CLEC1 ITAM receptor may be replaced by the FCERG cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CLEC1 ITAM receptor may be replaced by the FCGR2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CLEC1 ITAM receptor may be replaced by the GPVIA cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CLEC1 ITAM receptor may be replaced by the LILRB2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CLEC1 ITAM receptor may be replaced by the PECAM1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CLEC1 ITAM receptor may be replaced by the TLT1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CLEC1 ITAM receptor may be replaced by the CEACAM1 cellular domain or a part thereof to generate CPR.

[0169] For example, the EC domain of the FCGR2 ITAM receptor can be replaced by the CLEC1 EC domain or a part thereof to generate CPR. For example, the EC domain of the FCGR2 ITAM receptor can be replaced by the GPVIA EC domain or a part thereof to generate CPR. For example, the EC domain of the FCGR2 ITAM receptor can be replaced by the LILRB2 EC domain or a part thereof to generate CPR. For example, the EC domain of the FCGR2 ITAM receptor can be replaced by the PECAM1 EC domain or a part thereof to generate CPR. For example, the EC domain of the FCGR2 ITAM receptor can be replaced by the TLT1 EC domain or a part thereof to generate CPR. For example, the EC domain of the FCGR2 ITAM receptor can be replaced by the CEACAM1 EC domain or a part thereof to generate CPR.

[0170] For example, the TM domain of the FCGR2 ITAM receptor can be replaced by the FCERG TM domain or a part thereof to generate CPR. For example, the TM domain of the FCGR2 ITAM receptor can be replaced by the CLEC1 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCGR2 ITAM receptor can be replaced by the GPVIA TM domain or a part thereof to generate CPR. For example, the TM domain of the FCGR2 ITAM receptor can be replaced by the LILRB2 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCGR2 ITAM receptor can be replaced by the PECAM1 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCGR2 ITAM receptor can be replaced by the TLT1 TM domain or a part thereof to generate CPR. For example, the TM domain of the FCGR2 ITAM receptor can be replaced by the CEACAM1 TM domain or a part thereof to generate CPR.

[0171] For example, the signal peptide of the FCGR2 ITAM receptor may be replaced by the FCERG signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCGR2 ITAM receptor may be replaced by the CLEC1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCGR2 ITAM receptor may be replaced by the GPVIA signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCGR2 ITAM receptor may be replaced by the LILRB2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCGR2 ITAM receptor may be replaced by the PECAM1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCGR2 ITAM receptor may be replaced by the TLT1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the FCGR2 ITAM receptor may be replaced by the CEACAM1 signal peptide or a part thereof to generate CPR.

[0172] For example, the cellular domain of the FCGR2 ITAM receptor may be replaced by the FCERG cellular domain or a part thereof to generate CPR. For example, the cytoplasmic domain of the FCGR2 ITAM receptor may be replaced by the CLEC1 cytoplasmic domain or a part thereof to generate CPR. For example, the cellular domain of the FCGR2 ITAM receptor may be replaced by the GPVIA cellular domain or a part thereof to generate CPR. For example, the cellular domain of the FCGR2 ITAM receptor may be replaced by the LILRB2 cellular domain or a part thereof to generate CPR. For example, the cytoplasmic domain of the FCGR2 ITAM receptor may be replaced by the PECAM1 cytoplasmic domain or a part thereof to generate CPR. For example, the cytoplasmic domain of the FCGR2 ITAM receptor may be replaced by the TLT1 cytoplasmic domain or a part thereof to generate CPR. For example, the cellular domain of the FCGR2 ITAM receptor may be replaced by the CEACAM1 cellular domain or a part thereof to generate CPR.

[0173] For example, the EC domain of the GPVIA ITAM receptor can be replaced by the CLEC1 EC domain or a part thereof to generate CPR. For example, the EC domain of the GPVIA ITAM receptor can be replaced by the FCGR2 EC domain or a part thereof to generate CPR. For example, the EC domain of the GPVIA ITAM receptor can be replaced by the LILRB2 EC domain or a part thereof to generate CPR. For example, the EC domain of the GPVIA ITAM receptor can be replaced by the PECAM1 EC domain or a part thereof to generate CPR. For example, the EC domain of the GPVIA ITAM receptor can be replaced by the TLT1 EC domain or a part thereof to generate CPR. For example, the EC domain of the GPVIA ITAM receptor can be replaced by the CEACAM1 EC domain or a part thereof to generate CPR.

[0174] For example, the TM domain of the GPVIA ITAM receptor can be replaced by the FCERG TM domain or a part thereof to generate CPR. For example, the TM domain of the GPVIA ITAM receptor can be replaced by the CLEC1 TM domain or a part thereof to generate CPR. For example, the TM domain of the GPVIA ITAM receptor can be replaced by the FCGR2 TM domain or a part thereof to generate CPR. For example, the TM domain of the GPVIA ITAM receptor can be replaced by the LILRB2 TM domain or a part thereof to generate CPR. For example, the TM domain of the GPVIA ITAM receptor can be replaced by the PECAM1 TM domain or a part thereof to generate CPR. For example, the TM domain of the GPVIA ITAM receptor can be replaced by the TLT1 TM domain or a part thereof to generate CPR. For example, the TM domain of the GPVIA ITAM receptor can be replaced by the CEACAM1 TM domain or a part thereof to generate CPR.

[0175] For example, the signal peptide of the GPVIA ITAM receptor may be replaced by the FCERG signal peptide or a part thereof to generate CPR. For example, the signal peptide of the GPVIA ITAM receptor may be replaced by the CLEC1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the GPVIA ITAM receptor may be replaced by the FCGR2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the GPVIA ITAM receptor may be replaced by the LILRB2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the GPVIA ITAM receptor may be replaced by the PECAM1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the GPVIA ITAM receptor may be replaced by the TLT1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the GPVIA ITAM receptor may be replaced by the CEACAM1 signal peptide or a part thereof to generate CPR.

[0176] For example, the cellular domain of the GPVIA ITAM receptor may be replaced by the FCERG cellular domain or a part thereof to generate CPR. For example, the cellular domain of the GPVIA ITAM receptor may be replaced by the CLEC1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the GPVIA ITAM receptor may be replaced by the FCGR2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the GPVIA ITAM receptor may be replaced by the LILRB2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the GPVIA ITAM receptor may be replaced by the PECAM1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the GPVIA ITAM receptor may be replaced by the TLT1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the GPVIA ITAM receptor may be replaced by the CEACAM1 cellular domain or a part thereof to generate CPR.

[0177] In one embodiment, the EC domain of the ITIM receptor shown in Table 5 may be replaced with a domain from another ITAM and / or ITIM receptor.

[0178] For example, the EC domain of the LILRB2 ITIM receptor can be replaced by the CLEC1 EC domain or a part thereof to generate CPR. For example, the EC domain of the LILRB2 ITIM receptor can be replaced by the GPVIA EC domain or a part thereof to generate CPR. For example, the EC domain of the LILRB2 ITIM receptor can be replaced by the FCGR2 EC domain or a part thereof to generate CPR. For example, the EC domain of the LILRB2 ITIM receptor can be replaced by the PECAM1 EC domain or a part thereof to generate CPR. For example, the EC domain of the LILRB2 ITIM receptor can be replaced by the TLT1 EC domain or a part thereof to generate CPR. For example, the EC domain of the LILRB2 ITIM receptor can be replaced by the CEACAM1 EC domain or a part thereof to generate CPR.

[0179] For example, the TM domain of the LILRB2 ITIM receptor can be replaced by the FCERG TM domain or a part thereof to generate CPR. For example, the TM domain of the LILRB2 ITIM receptor can be replaced by the CLEC1 TM domain or a part thereof to generate CPR. For example, the TM domain of the LILRB2 ITIM receptor can be replaced by the GPVIA TM domain or a part thereof to generate CPR. For example, the TM domain of the LILRB2 ITIM receptor can be replaced by the FCGR2 TM domain or a part thereof to generate CPR. For example, the TM domain of the LILRB2 ITIM receptor can be replaced by the PECAM1 TM domain or a part thereof to generate CPR. For example, the TM domain of the LILRB2 ITIM receptor can be replaced by the TLT1 TM domain or a part thereof to generate CPR. For example, the TM domain of the LILRB2 ITIM receptor can be replaced by the CEACAM1 TM domain or a part thereof to generate CPR.

[0180] For example, the signal peptide of the LILRB2 ITIM receptor can be replaced by the FCERG signal peptide or a part thereof to generate CPR. For example, the signal peptide of the LILRB2 ITIM receptor can be replaced by the CLEC1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the LILRB2 ITIM receptor can be replaced by the GPVIA signal peptide or a part thereof to generate CPR. For example, the signal peptide of the LILRB2 ITIM receptor can be replaced by the FCGR2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the LILRB2 ITIM receptor can be replaced by the PECAM1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the LILRB2 ITIM receptor can be replaced by the TLT1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the LILRB2 ITIM receptor can be replaced by the CEACAM1 signal peptide or a part thereof to generate CPR.

[0181] For example, the cellular domain of the LILRB2 ITIM receptor can be replaced by the FCERG cellular domain or a part thereof to generate CPR. For example, the cellular domain of the LILRB2 ITIM receptor can be replaced by the CLEC1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the LILRB2 ITIM receptor can be replaced by the GPVIA cellular domain or a part thereof to generate CPR. For example, the cellular domain of the LILRB2 ITIM receptor can be replaced by the FCGR2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the LILRB2 ITIM receptor can be replaced by the PECAM1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the LILRB2 ITIM receptor can be replaced by the TLT1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the LILRB2 ITIM receptor can be replaced by the CEACAM1 cellular domain or a part thereof to generate CPR. For example, the EC domain of the PECAM1 ITIM receptor can be replaced by the CLEC1 EC domain or a part thereof to generate CPR. For example, the EC domain of the PECAM1 ITIM receptor can be replaced by the GPVIA EC domain or a part thereof to generate CPR. For example, the EC domain of the PECAM1 ITIM receptor can be replaced by the FCGR2 EC domain or a part thereof to generate CPR. For example, the EC domain of the PECAM1 ITIM receptor can be replaced by the LILRB2 EC domain or a part thereof to generate CPR. For example, the EC domain of the PECAM1 ITIM receptor can be replaced by the TLT1 EC domain or a part thereof to generate CPR. For example, the EC domain of the PECAM1 ITIM receptor can be replaced by the CEACAM1 EC domain or a part thereof to generate CPR.

[0182] For example, the TM domain of the PECAM1 ITIM receptor can be replaced by the FCERG TM domain or a part thereof to generate CPR. For example, the TM domain of the PECAM1 ITIM receptor can be replaced by the CLEC1 TM domain or a part thereof to generate CPR. For example, the TM domain of the PECAM1 ITIM receptor can be replaced by the GPVIA TM domain or a part thereof to generate CPR. For example, the TM domain of the PECAM1 ITIM receptor can be replaced by the FCGR2 TM domain or a part thereof to generate CPR. For example, the TM domain of the PECAM1 ITIM receptor can be replaced by the LILRB2 TM domain or a part thereof to generate CPR. For example, the TM domain of the PECAM1 ITIM receptor can be replaced by the TLT1 TM domain or a part thereof to generate CPR. For example, the TM domain of the PECAM1 ITIM receptor can be replaced by the CEACAM1 TM domain or a part thereof to generate CPR.

[0183] For example, the signal peptide of the PECAM1 ITIM receptor can be replaced by the FCERG signal peptide or a part thereof to generate CPR. For example, the signal peptide of the PECAM1 ITIM receptor can be replaced by the CLEC1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the PECAM1 ITIM receptor can be replaced by the GPVIA signal peptide or a part thereof to generate CPR. For example, the signal peptide of the PECAM1 ITIM receptor can be replaced by the FCGR2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the PECAM1 ITIM receptor can be replaced by the LILRB2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the PECAM1 ITIM receptor can be replaced by the TLT1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the PECAM1 ITIM receptor can be replaced by the CEACAM1 signal peptide or a part thereof to generate CPR.

[0184] For example, the cellular domain of the PECAM1 ITIM receptor can be replaced by the FCERG cellular domain or a part thereof to generate CPR. For example, the cellular domain of the PECAM1 ITIM receptor can be replaced by the CLEC1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the PECAM1 ITIM receptor can be replaced by the GPVIA cellular domain or a part thereof to generate CPR. For example, the cellular domain of the PECAM1 ITIM receptor can be replaced by the FCGR2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the PECAM1 ITIM receptor can be replaced by the LILRB2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the PECAM1 ITIM receptor can be replaced by the TLT1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CEACAM1 ITIM receptor can be replaced by the CEACAM1 cellular domain or a part thereof to generate CPR.

[0185] For example, the EC domain of the CEACAM1 ITIM receptor can be replaced by the CLEC1 EC domain or a part thereof to generate CPR. For example, the EC domain of the CEACAM1 ITIM receptor can be replaced by the GPVIA EC domain or a part thereof to generate CPR. For example, the EC domain of the CEACAM1 ITIM receptor can be replaced by the FCGR2 EC domain or a part thereof to generate CPR. For example, the EC domain of the CEACAM1 ITIM receptor can be replaced by the LILRB2 EC domain or a part thereof to generate CPR. For example, the EC domain of the CEACAM1 ITIM receptor can be replaced by the PECAM1 EC domain or a part thereof to generate CPR. For example, the EC domain of the CEACAM1 ITIM receptor can be replaced by the TLT1 EC domain or a part thereof to generate CPR.

[0186] For example, the TM domain of the CEACAM1 ITIM receptor can be replaced by the FCERG TM domain or a part thereof to generate CPR. For example, the TM domain of the CEACAM1 ITIM receptor can be replaced by the CLEC1 TM domain or a part thereof to generate CPR. For example, the TM domain of the CEACAM1 ITIM receptor can be replaced by the GPVIA TM domain or a part thereof to generate CPR. For example, the TM domain of the CEACAM1 ITIM receptor can be replaced by the FCGR2 TM domain or a part thereof to generate CPR. For example, the TM domain of the CEACAM1 ITIM receptor can be replaced by the LILRB2 TM domain or a part thereof to generate CPR. For example, the TM domain of the CEACAM1 ITIM receptor can be replaced by the PECAM1 TM domain or a part thereof to generate CPR. For example, the TM domain of the CEACAM1 ITIM receptor can be replaced by the TLT1 TM domain or a part thereof to generate CPR.

[0187] For example, the signal peptide of the CEACAM1 ITIM receptor can be replaced by the FCERG signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CEACAM1 ITIM receptor can be replaced by the CLEC1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CEACAM1 ITIM receptor can be replaced by the GPVIA signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CEACAM1 ITIM receptor can be replaced by the FCGR2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CEACAM1 ITIM receptor can be replaced by the LILRB2 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CEACAM1 ITIM receptor can be replaced by the PECAM1 signal peptide or a part thereof to generate CPR. For example, the signal peptide of the CEACAM1 ITIM receptor can be replaced by the TLT1 signal peptide or a part thereof to generate CPR.

[0188] For example, the cellular domain of the CEACAM1 ITIM receptor may be replaced by the FCERG cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CEACAM1 ITIM receptor may be replaced by the CLEC1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CEACAM1 ITIM receptor may be replaced by the GPVIA cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CEACAM1 ITIM receptor may be replaced by the FCGR2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CEACAM1 ITIM receptor may be replaced by the LILRB2 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CEACAM1 ITIM receptor may be replaced by the PECAM1 cellular domain or a part thereof to generate CPR. For example, the cellular domain of the CEACAM1 ITIM receptor may be replaced by the TLT1 cellular domain or a part thereof to generate CPR.

[0189] In some embodiments, the signal peptide, EC domain, TM domain, or cellular domain of the ITIM or ITAM receptor may be replaced by a portion of a domain from a different receptor. This portion may have a length in the range of 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or 20-30 nucleotides. This portion may be 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides of any of the sequences in Tables 6-10 of this specification, for example, SEQ ID NOs. 1-55. In a particular embodiment, the EC domain from the GPVIA ITAM receptor is Sequence ID No. 55.

[0190] 2. Antibodies or their fragments A CPR may contain a region with high specificity and affinity that includes any ligand or fragment or epitope that recognizes and binds to a target, such as a novel antigen on tumor cells. The region of the CPR may be a recognition component such as an antibody or variant thereof, or a variant or fragment thereof, that can bind to a target on tumor cells, such as a tumor-specific antigen (TSA), or a cytokine receptor, or a linked cytokine that leads to the recognition of a target cell having a native ligand or fragment of the receptor. In some embodiments, the CPR of the manipulated platelets described herein may be scFv. Antibodies, variants, or fragments thereof can be generated using conventional recombinant DNA techniques known in the art.

[0191] In some embodiments, the manipulated platelet includes a CPR having a region containing an antibody or antibody fragment for binding to a target of interest. For example, the CPR may include a variable heavy chain domain of an antibody. For example, the CPR may include a variable light chain domain of an antibody. Alternatively, the CPR of the manipulated platelet may include a kappa light chain or a fragment thereof that targets CD19.

[0192] In some embodiments, the antibody or antibody fragment is selected from Table 11. The antibodies are listed along with their DrugBank identifiers (DB IDs). [Table 11-1] [Table 11-2] [Table 11-3] [Table 11-4] [Table 11-5] Table 11-6 Table 11-7 Table 11-8 Table 11-9 Table 11-10

[0193] In some embodiments, antibodies may target or bind to antigens associated with a disease, disorder, or condition. Furthermore, antibodies or antibody fragments may be effective in treating a disease, disorder, or condition by binding to a target antigen. In some embodiments, the target of an antibody may be used to identify or describe the antibody. For example, 3F8 targets GD2 ganglioside for the treatment of neuroblastoma. For example, 8H9 targets B7-H3 for the treatment of neuroblastoma, sarcoma, and metastatic brain cancer. For example, avagovomab targets CA-125 (mimicking) for the treatment of ovarian cancer. For example, absiximab targets CD41 (integrin α-IIb) for the treatment of platelet aggregation inhibitors. For example, abituzumab targets CD51 for the treatment of cancer. For example, abrezekimab targets interleukin 13. For example, abrilumab targets integrin α4β7 for the treatment of inflammatory bowel disease, ulcerative colitis, and Crohn's disease. For example, actoxumab targets Clostridium difficile for the treatment of Clostridium difficile colitis. For example, adalimumab targets TNF-α for the treatment of rheumatoid arthritis, Crohn's disease, psoriasis vulgaris, psoriatic arthritis, ankylosing spondylitis, juvenile idiopathic arthritis, and neonatal hemolytic disease. For example, adekatumumab targets EpCAM for the treatment of prostate cancer and breast cancer. For example, atidorutoxumab targets Staphylococcus aureus alpha toxin. For example, aducanumab targets beta-amyloid for the treatment of Alzheimer's disease. For example, afacebikumab targets IL17A and IL17F for the treatment of multiple sclerosis. For example, aferimomab targets TNF-α for the treatment of sepsis. For example, aracizumab pego targets VEGFR2 for the treatment of cancer. For example, alemtuzumab targets CD52 for the treatment of multiple sclerosis. For example, alirocumab targets PCSK9 for the treatment of hypercholesterolemia. For example, artumomab pentetate targets CEA for the treatment (diagnosis) of colorectal cancer. For example, amatuximab targets mesoserine for the treatment of cancer.For example, anatumomab mafenatox targets TAG-72 for the treatment of non-small cell lung cancer. For example, andecaliximab targets gelatinase B for the treatment of gastric cancer or gastroesophageal junction adenocarcinoma. For example, anetumablavutansine targets MSLN for the treatment of cancer. For example, aniflorumab targets interferon α / β receptors for the treatment of systemic lupus erythematosus. For example, anlukinzumab targets IL-13 for the treatment of asthma. For example, apolizumab targets HLA-DR for the treatment of hematological malignancies. For example, appletumab ixadotin targets FGFR2. For example, alsitumomab targets CEA for the treatment (diagnosis) of gastrointestinal cancers. For example, askrinbakumab targets activin receptor-like kinase 1 for the treatment of cancer. For example, aselizumab targets L-selectin (CD62L) to treat severely injured patients. For example, atezolizumab targets PD-L1 to treat cancer. For example, atinumab targets RTN4. For example, atromimubab targets rhesus factor to treat neonatal hemolytic disease [citation needed]. For example, avelumab targets PD-L1 to treat cancer. For example, adintuxizumab vedotin targets CD319 to treat cancer. For example, bapinuzumab targets beta-amyloid to treat Alzheimer's disease. For example, basiliximab targets CD25 (α chain of the IL-2 receptor) to treat the prevention of organ transplant rejection. For example, bavituximab targets phosphatidylserine to treat cancer and viral infections. For example, BCD-100 targets PD-1 for the treatment of melanoma. For example, vectumomab targets CD22 for the treatment (detection) of non-Hodgkin lymphoma. For example, vegeromab targets DPP4. For example, belantamab mafodotin targets BCMA for the treatment of cancer. For example, belimumab targets BAFF for the treatment of non-Hodgkin lymphoma. For example, bemarituzumab targets FGFR2 for the treatment of gastric cancer or gastroesophageal junction adenocarcinoma. For example, benralizumab targets CD125 for the treatment of asthma.For example, berlimatoxumab targets leucocidine, a two-component of Staphylococcus aureus. For example, bermekimab targets IL1A for the treatment of colorectal cancer. For example, bersanlimab targets ICAM-1. For example, bertilimumab targets CCL11 (eotaxin-1) for the treatment of severe allergic diseases. For example, becylesomab targets CEA-related antigens for the treatment (detection) of inflammatory lesions and metastases. For example, bevacizumab targets VEGF-A for the treatment of metastatic cancer and retinopathy of prematurity. For example, bezlotoxumab targets Clostridium difficile for the treatment of Clostridium difficile colitis. For example, bisilomab targets fibrin II, a beta-chain, for the treatment (diagnosis) of thromboembolism. For example, bimaglumab targets ACVR2B for the treatment of myostatin inhibitors. For example, bimekizumab targets IL17A and IL17F for the treatment of ankylosing spondylitis and psoriasis. For example, virtamimab targets serum amyloid A protein for the treatment of amyloidosis. For example, vibatuzumab meltansine targets CD44v6 for the treatment of squamous cell carcinoma. For example, preserumab targets CD40 for the treatment of organ transplant rejection. For example, blinatumomab targets CD19 for the treatment of pre-B ALL (CD19+). For example, bronzvetomab targets CD20. For example, brosozumab targets SOST for the treatment of osteoporosis. For example, vococizumab targets neuronal apoptosis-regulating proteinase 1 for the treatment of dyslipidemia. For example, brazicumab targets IL23 for the treatment of Crohn's disease. For example, brentuximab vedotin targets CD30 (TNFRSF8) for the treatment of Hodgkin lymphoma and anaplastic large cell lymphoma. For example, briakinumab targets IL-12 and IL-23 for the treatment of psoriasis, rheumatoid arthritis, inflammatory bowel disease, and multiple sclerosis. For example, brodalumab targets IL-17 for the treatment of plaque psoriasis. For example, brolucizumab targets VEGFA for the treatment of wet age-related macular degeneration.For example, brontuctuzumab targets Notch1 for the treatment of cancer. For example, burosumab targets FGF23 for the treatment of X-linked hypophosphatemia. For example, kabilizumab targets CSF1R for the treatment of metastatic pancreatic cancer. For example, camidanrumab tesirin targets CD25 for the treatment of B-cell Hodgkin lymphoma, non-Hodgkin lymphoma, acute lymphoblastic leukemia, and acute myeloid leukemia. For example, camrelizumab targets PD-1 for the treatment of hepatocellular carcinoma. For example, canakinumab targets IL-1 for the treatment of cryopyrin-associated cyclical syndrome. For example, cantuzumab meltansine targets mucin CanAg for the treatment of colorectal cancer. For example, cantuzumab meltansine targets MUC1 for the treatment of cancer. For example, caplacizumab targets VWF for the treatment of thrombotic thrombocytopenic purpura and thrombosis. For example, capromab pendetide targets prostate cancer cells for the treatment (detection) of prostate cancer. For example, carlumab targets MCP-1 for the treatment of oncological / immunological indications. For example, carotuximab targets endoglin for the treatment of angiosarcoma. For example, catumakisomab targets EpCAM and CD3 for the treatment of ovarian cancer, malignant ascites, and gastric cancer. For example, the cbr96-doxorubicin immune complex targets Lewis Y antigen for the treatment of cancer. For example, sedelizumab targets CD4 for the prevention of organ transplant rejection and the treatment of autoimmune diseases. For example, semiprimab targets PCDC1 for the treatment of cutaneous squamous cell carcinoma. For example, sergutuzumab amnaleukin targets IL2 for the treatment of cancer. For example, certolizumab pegol targets TNF-α for the treatment of Crohn's disease, rheumatoid arthritis, axial spondyloarthritis, and psoriatic arthritis. For example, cetrelimab targets PD-1 for the treatment of cancer. For example, cetuximab targets EGFR for the treatment of metastatic colorectal cancer and head and neck cancer. For example, cibisatamab targets CEACAM5 for the treatment of cancer. For example, sirumutuzumab targets ROR1 for the treatment of chronic lymphocytic leukemia.For example, sitatuzumab vodatox targets EpCAM for the treatment of ovarian cancer and other solid tumors. For example, xixtumumab targets the IGF-1 receptor (CD221) for the treatment of solid tumors. For example, crazakizumab targets IL6 for the treatment of rheumatoid arthritis. For example, clenoliximab targets CD4 for the treatment of rheumatoid arthritis. For example, cribatuzumab tetraxetan targets MUC1 for the treatment of pancreatic cancer. For example, codlituzumab targets glypican 3 for the treatment of cancer. For example, cofetuzumab peridotin targets PTK7 for the treatment of cancer. For example, coltuximab brabutansine targets CD19 for the treatment of cancer. For example, conatumumab targets TRAIL-R2 for the treatment of cancer. For example, concizumab targets TFPI for the treatment of bleeding. For example, cosflobiximab targets the Ebola virus glycoprotein for the treatment of the Ebola virus. For example, crenezumab targets 1-40-β-amyloid for the treatment of Alzheimer's disease. For example, chryzanlizumab targets selectin P for the treatment of sickle cell disease. For example, clotedumab targets GCGR for the treatment of diabetes. For example, CR6261 targets influenza A hemagglutinin for the treatment of infections / influenza A. For example, xatuzumab targets CD70 for the treatment of cancer. For example, dasetuzumab targets CD40 for the treatment of hematological malignancies. For example, daclizumab targets CD25 (the α chain of the IL-2 receptor) for the prevention of organ transplant rejection and multiple sclerosis. For example, dalotuzumab targets the IGF-1 receptor (CD221) for the treatment of cancer. For example, dapyrolizumab pegol targets CD154 (CD40L). For example, daratumumab targets CD38 for the treatment of multiple myeloma. For example, dectrecumab targets IL-13. For example, demcizumab targets DLL4 for the treatment of cancer. For example, denintuzumab mahodotin targets CD19 for the treatment of cancer. For example, denosumab targets RANKL for the treatment of osteoporosis and bone metastases.For example, depatuxizumab mahodotin targets EGFR for the treatment of glioblastoma. For example, delrotuximab biotin targets histone complexes for the treatment of relapsed glioblastoma multiforme. For example, detumomab targets B lymphoma cells for the treatment of lymphoma. For example, dezamizumab targets serum amyloid-P component. For example, dinutuximab targets GD2 ganglioside for the treatment of neuroblastoma. For example, dilidabumab targets influenza. For the treatment of Enza A, hemagglutinins are targeted. For example, domaglozumab targets GDF-8 for the treatment of Duchenne muscular dystrophy. For example, dostallimab targets PCDP1 for the treatment of cancer. For example, drodizumab targets DR5 for the treatment of cancer. For example, DS-8201 targets HER2 for the treatment of gastric or gastroesophageal junction adenocarcinoma. For example, duligotuzumab targets ERBB3 (HER3) for the treatment of testicular cancer. For example, dupilumab targets IL4 for the treatment of atopic diseases. For example, durvalumab targets PD-L1 for the treatment of cancer. For example, dusigituzumab targets ILGF2 for the treatment of B-cell malignancies. For example, duvortuxizumab targets CD19 and CD3E for the treatment of cancer. For example, eclomeximab targets GD3 ganglioside for the treatment of malignant melanoma. For example, eculizumab targets C5 for the treatment of paroxysmal nocturnal hemoglobinuria and atypical HUS. For example, edovacomab targets endotoxin for the treatment of sepsis caused by Gram-negative bacteria. For example, edrecolomab targets EpCAM for the treatment of colorectal cancer. For example, efalizumab targets LFA-1 (CD11a) for the treatment of psoriasis (by inhibiting T cell migration). For example, efungumab targets Hsp90 for the treatment of invasive Candida infection. For example, elderumab targets interferon-gamma-inducible protein for the treatment of Crohn's disease and ulcerative colitis. For example, elezanumab targets RGMA for the treatment of spinal cord injury and multiple sclerosis. For example, elgemzumab targets ERBB3 (HER3) for the treatment of cancer. For example, elotuzumab targets SLAMF7 for the treatment of multiple myeloma. For example, elcilimomab targets IL-6. For example, emuctuzumab targets CSF1R for the treatment of cancer. For example, emaparmab targets interferon-gamma for the treatment of hemophagocytic lymphohistiocytosis. For example, emibetuzumab targets HHGFR for the treatment of cancer.For example, emicizumab targets activated F9 and F10 for the treatment of hemophilia A. For example, enapotamab vedotin targets AXL for the treatment of cancer. For example, enabatuzumab targets the TWEAK receptor for the treatment of cancer and other conditions. For example, enfortumab vedotin targets nectin-4 for the treatment of urothelial carcinoma. For example, enrimomab pegol targets ICAM-1 (CD54). For example, enobrituzumab targets CD276 for the treatment of cancer. For example, enokizumab targets IL9 for the treatment of asthma. For example, enoticumab targets DLL4. For example, encituximab targets 5AC for the treatment of cancer. For example, epitumomab citucetan targets epicyalin. For example, epratuzumab targets CD22 for the treatment of cancer and SLE. For example, eptinezumab targets calcitonin gene-related peptide for the treatment of migraines. For example, erenumab targets CGRP for the treatment of migraines. For example, erulizumab targets ITGB2 (CD18) for the treatment of heart attacks, strokes, and traumatic shock. For example, ertzumaxomab targets HER2 / neu and CD3 for the treatment of breast cancer, etc. For example, etalacizumab targets integrin αvβ3 for the treatment of melanoma, prostate cancer, ovarian cancer, etc. For example, etidyrimab targets TIGIT. For example, etrolizumab targets integrin β7 for the treatment of inflammatory bowel disease. For example, evinacumab targets angiopoietin 3 for the treatment of dyslipidemia. For example, evolocumab targets PCSK9 for the treatment of hypercholesterolemia. For example, exvivirumab targets the hepatitis B surface antigen for the treatment of hepatitis B. For example, fanolesomab targets CD15 for the treatment (diagnosis) of appendicitis. For example, falalimomab targets the interferon receptor. For example, falisimab targets VEGF-A and Ang-2 for the treatment of angiogenesis and ophthalmic vascular disease. For example, farletuzumab targets folate receptor 1 for the treatment of ovarian cancer. For example, facinumab targets HNGF for the treatment of acute sciatica.For example, FBTA05 targets CD20 for the treatment of chronic lymphocytic leukemia. For example, felbizumab targets respiratory syncytial virus for the treatment of respiratory syncytial virus infection. For example, fezakinumab targets IL-22 for the treatment of rheumatoid arthritis and psoriasis. For example, fibatuzumab targets ephrin receptor A3. For example, ficratuzumab targets HGF for the treatment of cancer. For example, figitumumab targets the IGF-1 receptor (CD221) for the treatment of adrenocortical carcinoma and non-small cell lung cancer. For example, filibumab targets hemagglutinin of influenza A virus. For example, frambotumab targets TYRP1 (glycoprotein 75) for the treatment of melanoma. For example, fretikumab targets IL20 for the treatment of rheumatoid arthritis. For example, flotetuzumab targets the IL3 receptor for the treatment of hematological malignancies. For example, fontlizumab targets IFN-γ for the treatment of Crohn's disease. For example, folarumab targets CD3 epsilon. For example, folavirumab targets rabies virus glycoprotein for the treatment (prevention) of rabies. For example, fremanezumab targets calcitonin gene-related peptide alpha for the treatment of migraines and cluster headaches. For example, fresolimmab targets TGF-β for the treatment of idiopathic pulmonary fibrosis, focal segmental glomerulosclerosis, and cancer. For example, flobokimab targets PCSK9 for the treatment of hypercholesterolemia. For example, flunebetomab targets NGF. For example, fluranumab targets NGF for the treatment of pain. For example, futuximab targets EGFR for the treatment of cancer. For example, galcanezumab targets calcitonin to treat migraines. For example, galiximab targets CD80 to treat B-cell lymphoma. For example, gancotamab targets HER2 / neu to treat cancer. For example, ganitumab targets the IGF-1 receptor (CD221) to treat cancer. For example, gantenerumab targets beta-amyloid to treat Alzheimer's disease. For example, gatipotuzumab targets MUC1 to treat cancer.For example, gabirimomab targets CD147 (basidine) to treat graft-versus-host disease. For example, gesibumab targets hemagglutinin HA. For example, gemtuzumab ozogamicin targets CD33 to treat acute myeloid leukemia. For example, gevokizumab targets IL-1β to treat diabetes mellitus. For example, zilvetomab targets PCDC1. For example, dimucirumab targets CSF2 to treat rheumatoid arthritis. For example, girentuximab targets carbonic anhydrase 9 (CA-IX) to treat clear cell renal cell carcinoma. For example, glembatumumab vedotin targets GPNMB to treat melanoma and breast cancer. For example, golimumab targets TNF-α to treat rheumatoid arthritis, psoriatic arthritis, and ankylosing spondylitis. For example, gomiliximab targets CD23 (IgE receptor) for the treatment of allergic asthma. For example, gosulanemab targets tau protein for the treatment of progressive supranuclear palsy. For example, guselkumab targets IL23 for the treatment of psoriasis. For example, ianarumab targets BAFF-R for the treatment of autoimmune hepatitis. For example, ibalizumab targets CD4 for the treatment of HIV infection. For example, IBI308 targets PD-1 for the treatment of squamous cell non-small cell lung cancer. For example, ibritumomab tiuxetan targets CD20 for the treatment of non-Hodgkin lymphoma. For example, ikulkumab targets VEGFR-1 for the treatment of cancer. For example, idarucizumab targets dabigatran for the treatment of dabigatran anticoagulant reversal. For example, ifabotuzumab targets EPHA3. For example, igovomab targets CA-125 for the treatment (diagnosis) of ovarian cancer. For example, iradatuzumab vedotin targets CD97B for cancer treatment. For example, IMAB362 targets CLDN18.2 for the treatment of gastrointestinal adenocarcinoma and pancreatic tumors. For example, imarumab targets MIF for cancer treatment. For example, imaprelimab targets MCAM. For example, imisilomab targets cardiac myosin for cardiac imaging treatment. For example, imugatuzumab targets EGFR for cancer treatment.For example, incrumab targets selectin P for the treatment of cardiovascular disease. For example, indatuximab tansine targets SDC1 for the treatment of cancer. For example, indusatumab vedotin targets GUCY2C for the treatment of cancer. For example, inebilizumab targets CD19 for the treatment of cancer, systemic sclerosis, and multiple sclerosis. For example, infliximab targets TNF-α for the treatment of rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, psoriasis, Crohn's disease, and ulcerative colitis. For example, intetumumab targets CD51 for the treatment of solid tumors (prostate cancer, melanoma). For example, inorimomab targets CD25 (the α chain of the IL-2 receptor) for the treatment of graft-versus-host disease. For example, inotuzumab ozogamicin targets CD22 for the treatment of ALL. For example, ipilimumab targets CD152 for the treatment of melanoma. For example, Iomab-B targets CD45 for the treatment of bone marrow resection. For example, iratumumab targets CD30 (TNFRSF8) for the treatment of Hodgkin lymphoma. For example, isatuximab targets CD38 for the treatment of multiple myeloma. For example, iscalimab targets CD40. For example, istilatumab targets IGF1R, CD221 for the treatment of advanced solid tumors. For example, itolizumab targets CD6 for the treatment of psoriasis. For example, ixekizumab targets IL17A for the treatment of autoimmune diseases. For example, keriximab targets CD4 for the treatment of chronic asthma. For example, rabetuzumab targets CEA for the treatment of colorectal cancer. For example, lacunotuzumab targets CSF1 and MCSF for the treatment of cancer. For example, radilatuzumab vedotin targets LIV-1 for the treatment of cancer. For example, lampalizumab targets CFD for the treatment of geographical atrophy secondary to age-related macular degeneration. For example, lanadermab targets kallikrein for the treatment of angioedema. For example, landgrozumab targets GDF-8 for the treatment of muscle wasting disease. For example, laprituximab emtansine targets EGFR.For example, ralkabiximab targets the Ebola virus glycoprotein for the treatment of the Ebola virus. For example, lebrikizumab targets IL-13 for the treatment of asthma. For example, remaresomab targets NCA-90 (granulocyte antigen) for the treatment of diagnostic agents. For example, lendarizumab targets C5. For example, lembervimab targets the hepatitis B surface antigen for the treatment of hepatitis B. For example, rengirumab targets chronic myelomonocytic leukemia and juvenile myelomonocytic leukemia. For example, CSF2 is targeted to treat diseases. For instance, reldelimumab targets TGF beta-2 to treat the reduction of scarring after glaucoma surgery. For instance, relonlimab targets CCR5. For instance, resofabumab targets hemagglutinin HA. For instance, letrizumab targets TRAP for the treatment of inflammatory diseases. For instance, lexatumumab targets TRAIL-R2 for the treatment of cancer. For instance, rivivirumab targets hepatitis B surface antigen for the treatment of hepatitis B. For instance, rifastuzumab vedotin targets sodium phosphate cotransporter for the treatment of cancer. For instance, rigerizumab targets IGHE for the treatment of severe asthma and chronic spontaneous urticaria. For instance, roncustuximab tesirin targets CD19 for the treatment of cancer. For example, rosatuxizumab vedotin targets EGRF, ERBB1 HER1 for the treatment of cancer. For example, rilotomab satetraxetan targets CD37 for the treatment of cancer. For example, lintuzumab targets CD33 for the treatment of cancer. For example, lirirumab targets KIR2D for the treatment of solid tumors and hematological malignancies. For example, rodelcizumab targets PCSK9 for the treatment of hypercholesterolemia. For example, lokivetomab targets Canis lupus familiaris IL31 for the treatment of clinical symptoms of canine atopic dermatitis

[50] . For example, lorbotuzumab meltansine targets CD56 for the treatment of cancer. For example, lucatumumab targets CD40 for the treatment of multiple myeloma, non-Hodgkin lymphoma, and Hodgkin lymphoma. For example, rulizumab pegol targets CD28 for the treatment of autoimmune diseases. For example, lumiliquisimab targets CD23 (IgE receptor) for the treatment of chronic lymphocytic leukemia. For example, lumuretuzumab targets ERBB3 (HER3) for the treatment of cancer. For example, rupatumab amadotin targets LYPD3. For example, rituximab targets interleukin-1 alpha. For example, mapatumumab targets TRAIL-R1 for the treatment of cancer. For example, margetuximab targets HER2 for the treatment of breast cancer.For example, marstatimab targets TFPI for the treatment of bleeding associated with hemophilia. For example, maslimomab targets the T cell receptor. For example, maprilimumab targets the GMCSF receptor α chain for the treatment of rheumatoid arthritis. For example, matuzumab targets EGFR for the treatment of colorectal cancer, lung cancer, and gastric cancer. For example, mepolizumab targets IL-5 for the treatment of asthma and leukocyte disorders. For example, meterimumab targets TGF beta 1 for the treatment of systemic sclerosis. For example, milatuzumab targets CD74 for the treatment of multiple myeloma and other hematological malignancies. For example, minretumomab targets TAG-72 for tumor detection and treatment. For example, mirikizumab targets IL23A for the treatment of psoriasis. For example, milbetuximab sorabtansine targets the folate receptor alpha for the treatment of ovarian cancer. For example, mitumomab targets GD3 ganglioside for the treatment of small cell lung cancer. For example, modotuximab targets EGFR extracellular domain III for the treatment of cancer. For example, mogamulizumab targets CCR4 for the treatment of adult T-cell leukemia / lymphoma. For example, monalizumab targets NKG2A for the treatment of rheumatoid arthritis, gynecological malignancies, and other cancers. For example, morolimumab targets rhesus factor. For example, mosnetuzumab targets CD3E, MS4A1, and CD20 for the treatment of cancer. For example, motavizumab targets respiratory syncytial virus for the treatment (prevention) of respiratory syncytial virus. For example, moxetumomab pasudotox targets CD22 for the treatment of hairy cell leukemia. For example, muromonab-CD3 targets CD3 for the treatment of organ transplant rejection. For example, nacolomab butafenatox targets the C242 antigen for the treatment of colorectal cancer. For example, namilumab targets CSF2. For example, naptumomab estafenatox targets 5T4 for the treatment of non-small cell lung cancer and renal cell carcinoma. For example, naratuximab emtansine targets CD37 for treatment. For example, nalnatumab targets MST1R (also known as RON) for the treatment of cancer.For example, natalizumab targets integrin α4 for the treatment of multiple sclerosis and Crohn's disease. For example, nabixixizumab targets DLL4 and VEGFA for the treatment of cancer. For example, navibu-mab targets the hemagglutinin HA of the influenza A virus. For example, naxitamab targets c-Met for the treatment of high-risk neuroblastoma and refractory myeloid disease. For example, nevacumab targets endotoxin for the treatment of sepsis. For example, nesitumumab targets EGFR for the treatment of non-small cell lung cancer. For example, nemolizumab targets IL31RA for the treatment of eczema

[53] . For example, NEOD001 targets amyloid for the treatment of primary systemic amyloidosis. For example, nererimomab targets TNF-α. For example, nesbukumab targets angiopoietin 2 for the treatment of cancer. For example, netakimab targets interleukin-17A. For example, nimotuzumab targets EGFR for the treatment of squamous cell carcinoma, head and neck cancer, nasopharyngeal cancer, and glioma. For example, nircevimab targets RSVFR for the treatment of respiratory syncytial virus. For example, nivolumab targets PD-1 for the treatment of cancer. For example, nofetumomab merpentan treats (diagnoses) cancer. For example, obiltoxaximab targets Bacillus anthracis anthrax for the treatment of anthrax spores. For example, obinutuzumab targets CD20 for the treatment of chronic lymphocytic leukemia. For example, okalatuzumab targets CD20 for the treatment of cancer. For example, ocrelizumab targets CD20 for the treatment of rheumatoid arthritis and lupus erythematosus. For example, odulimomab targets LFA-1 (CD11a) for the treatment of organ transplant rejection and the prevention of immune diseases. For example, ofatumumab targets CD20 for the treatment of chronic lymphocytic leukemia. For example, olaratumab targets PDGF-Rα for the treatment of cancer. For example, oleculumab targets 5'-nucleotidase for the treatment of pancreatic cancer and colorectal cancer.For example, orendalizumab targets complement component C5a for the treatment of systemic lupus erythematosus, lupus nephritis, and acute graft-versus-host disease. For example, orokizumab targets IL6 for the treatment of rheumatoid arthritis. For example, omalizumab targets the IgE Fc region for the treatment of allergic asthma. For example, ombrutamab targets CD276 for the treatment of cancer. For example, OMS721 targets MASP-2 for the treatment of atypical hemolytic uremic syndrome. For example, onartuzumab targets human scattering factor receptor kinase for the treatment of cancer. For example, ontuxizumab targets TEM1 for the treatment of cancer. For example, onbatirimab targets VSIR. For example, opicinumab targets LINGO-1 for the treatment of multiple sclerosis. For example, oportuzumab monatox targets EpCAM for the treatment of bladder cancer. For example, olegobomab targets CA-125 for the treatment of ovarian cancer. For example, orticumab targets oxLDL. For example, otelixizumab targets CD3 for the treatment of type 1 diabetes. For example, ocilimab targets GMCSF for the treatment of osteoarthritis and rheumatoid arthritis. For example, otreltuzumab targets CD37 for the treatment of cancer. For example, oxelumab targets OX-40 for the treatment of asthma. For example, ozanezumab targets NOGO-A for the treatment of ALS and multiple sclerosis. For example, ozoralizumab targets TNF-α for the treatment of inflammation. For example, pagibaximab targets lipoteichoic acid for the treatment of sepsis (Staphylococcus). For example, palivizumab targets the F protein of respiratory syncytial virus for the treatment (prevention) of respiratory syncytial virus. For example, pamrebrumab targets CTGF for the treatment of idiopathic pulmonary fibrosis (IPF) and pancreatic cancer. For example, panitumumab targets EGFR for the treatment of colorectal cancer. For example, pancomab targets tumor-specific glycosylation of MUC1 for the treatment of ovarian cancer. For example, panobacumab targets Pseudomonas aeruginosa for the treatment of Pseudomonas aeruginosa infections. For example, pulsatuzumab targets EGFL7 for the treatment of cancer.For example, pascolizumab targets IL-4 for the treatment of asthma. For example, pasotuxizumab targets folate hydrolase for the treatment of cancer. For example, pateclizumab targets LTA for the treatment of TNF. For example, patrizumab targets ERBB3 (HER3) for the treatment of cancer. For example, PDR001 targets PD-1 for the treatment of melanoma. For example, pembrolizumab targets PD-1 for the treatment of melanoma and other cancers. For example, pemtumomab targets MUC1 for the treatment of cancer. For example, perakizumab targets IL17A for the treatment of arthritis. For example, pertuzumab targets HER2 / neu for the treatment of cancer. For example, pexerizumab targets C5 for the treatment of reducing side effects of cardiac surgery. For example, pizilizumab targets PD-1 for the treatment of cancer and infections. For example, pinatuzumab vedotin targets CD22 for cancer treatment. For example, pinitumomab targets adenocarcinoma antigens for the treatment (imaging) of adenocarcinoma. For example, pracumab targets human TNF for the treatment of pain and inflammatory diseases. For example, prozarizumab targets CCR2 for the treatment of diabetic nephropathy and arteriovenous graft patency. For example, pogalizumab targets TNFR superfamily member 4. For example, polatuzumab vedotin targets CD79B for the treatment of diffuse large B-cell lymphoma. For example, ponezumab targets human beta-amyloid for the treatment of Alzheimer's disease. For example, polgabiximab targets Zaire Ebola virus glycoprotein for the treatment of Ebola virus disease. For example, pracinezumab targets NACP for the treatment of Parkinson's disease. For example, prezarizumab targets ICOSL. For example, priliximab targets CD4 for the treatment of Crohn's disease and multiple sclerosis. For example, pritoxaximab targets E. coli shiga toxin type 1. For example, pritumumab targets vimentin for the treatment of brain cancer. For example, PRO140 targets CCR5 for the treatment of HIV infection.For example, quilizumab targets IGHE for the treatment of asthma. For example, lacosumomab targets NGNA gangliosides for the treatment of non-small cell lung cancer. For example, radrezumab targets fibronectin extradomain B for cancer treatment. For example, rafibirumab targets rabies virus glycoprotein for the treatment (prevention) of rabies. For example, larpancizumab targets neuronal apoptosis-regulating proteinase 1 for the treatment of dyslipidemia. For example, ramucirumab... For example, lanebetomab targets VEGFR2 for the treatment of solid tumors. For example, ranibizumab targets NGF for the treatment of canine osteoarthritis. For example, ranibizumab targets VEGF-A for the treatment of wet macular degeneration. For example, laxibakumab targets anthrax toxin and protective antigens for the treatment (prevention and treatment) of Bacillus anthrax. For example, lavagalimab targets CD40 for the treatment of Crohn's disease. For example, raburizumab targets C5 for the treatment of paroxysmal nocturnal hemoglobinuria and atypical hemolytic uremic syndrome. For example, refanezumab targets myelin-related glycoprotein for the treatment of motor function recovery after stroke. For example, regavirumab targets cytomegalovirus glycoprotein B for the treatment of cytomegalovirus infection. For example, relatrimab targets LAG3 for the treatment of melanoma. For example, remtolumab targets interleukin-17-alpha (TNF-α). For example, reslizumab targets IL-5 for the treatment of inflammation of the airways, skin, and gastrointestinal tract. For example, rilotumumab targets HGF for the treatment of solid tumors. For example, linucumab targets platelet-derived growth factor receptor beta for the treatment of angiogenic age-related macular degeneration. For example, risankizumab targets IL23A for the treatment of Crohn's disease, psoriasis, psoriatic arthritis, and asthma. For example, rituximab targets CD20 for the treatment of lymphoma, leukemia, and certain autoimmune diseases. For example, rivabazumab pegol targets the Pseudomonas aeruginosa type III secretory system. For example, lobatumumab targets the IGF-1 receptor (CD221) for cancer treatment. For example, Rmab targets rabies virus G glycoprotein for the treatment of post-exposure prophylaxis for rabies. For example, loredumab targets RHD for the treatment of Rh incompatibility. For example, romikimab targets interleukin-13. For example, romosozumab targets sclerostin for the treatment of osteoporosis. For example, lontalizumab targets IFN-α for the treatment of systemic lupus erythematosus. For example, rosmantuzumab targets root plate-specific spondin 3 for the treatment of cancer.For example, lovalpituzumab tecillin targets DLL3 for the treatment of small cell lung cancer. For example, lobelizumab targets CD11 and CD18 for the treatment of hemorrhagic shock. For example, rozanolixizumab targets FCGRT. For example, laplizumab targets CD154 (CD40L) for the treatment of rheumatic diseases. For example, SA237 targets IL-6R for the treatment of neuromyelitis optica and neuromyelitis optica spectrum disorder. For example, sacituzumab govitecan targets TROP-2 for the treatment of triple-negative breast cancer. For example, samalizumab targets CD200 for the treatment of cancer. For example, samlotamab vetotin targets LRRC15 for the treatment of cancer. For example, sarilumab targets IL-6 for the treatment of rheumatoid arthritis and ankylosing spondylitis. For example, satralizumab targets the IL6 receptor for the treatment of neuromyelitis optica. For example, satumomab pendetide targets TAG-72 for the treatment (diagnosis) of cancer. For example, secukinumab targets IL17A for the treatment of uveitis and rheumatoid arthritis psoriasis. For example, sericrelumab targets CD40. For example, serivanthumab targets ERBB3 (HER3) for the treatment of cancer. For example, cetoxaximab targets E. coli Shiga toxin type 2. For example, setorusumab targets SOST. For example, sevilumab targets cytomegalovirus for the treatment of cytomegalovirus infection. For example, cibrotuzumab targets FAP for the treatment of cancer. For example, SGN-CD19A targets CD19 for the treatment of acute lymphoblastic leukemia and B-cell non-Hodgkin lymphoma. For example, SHP647 targets mucosal adrenergic cell adhesion molecules for the treatment of Crohn's disease. For example, cifalimumab targets IFN-α for the treatment of SLE, dermatomyositis, and polymyositis. For example, siltuximab targets IL-6 for the treatment of cancer. For example, simtuzumab targets LOXL2 for the treatment of fibrosis. For example, ciprizumab targets CD2 for the treatment of psoriasis and graft-versus-host disease (prevention). For example, siltratumab vetotin targets SLITRK6 for the treatment of cancer.For example, silkumab targets IL-6 for the treatment of rheumatoid arthritis. For example, sofituzumab vedotin targets CA-125 for the treatment of ovarian cancer. For example, solanezumab targets beta-amyloid for the treatment of Alzheimer's disease. For example, solitomab targets EpCAM for the treatment of gastrointestinal, lung, and other cancers. For example, sonepcizumab targets sphingosine-1-phosphate for the treatment of choroidal and retinal neovascularization. For example, sontuzumab targets epicyalin. For example, spartalizumab targets PDCD1 and CD279 for the treatment of melanoma. For example, stamlumab targets myostatin for the treatment of muscular dystrophy. For example, thresomab targets NCA-90 (granulocyte antigen) for the treatment of osteomyelitis. For example, subtabumab targets RSVFR for the treatment of lower respiratory tract diseases requiring medical intervention. For example, stimulimab targets C1s for the treatment of cold agglutinin disease. For example, subizumab targets HIV-1 for the treatment of viral infections. For example, subutiratoximab targets Staphylococcus aureus alpha toxin for the treatment of hospital-acquired pneumonia. For example, tavarumab targets BAFF for the treatment of B-cell cancer. For example, tacuzumab tetraxetan targets alpha-fetoprotein for cancer treatment. For example, tadocizumab targets integrin αIIbβ3 for the treatment of percutaneous coronary intervention. For example, talakotsuzumab targets CD123. For example, talizumab targets IgE for the treatment of allergic reactions. For example, tamtuvetomab targets CD52 for treatment. For example, tanezumab targets NGF for the treatment of pain. For example, tapritumomab paptox targets CD19 for the treatment of cancer. For example, tarexuzumab targets the Notch receptor for the treatment of cancer. For example, tavolimab targets CD134 for the treatment of cancer. For example, tefivazumab targets agglutinating factor A for the treatment of Staphylococcus aureus infection. For example, terisotuzumab vedotin targets HGFR for the treatment of cancer.For example, tenatumomab targets tenascin C for the treatment of cancer. For example, teneriximab targets CD40 for the treatment of autoimmune diseases and the prevention of organ transplant rejection. For example, teprizumab targets CD3 for the treatment of type 1 diabetes. For example, tepoditamab targets dendritic cell-associated lectin 2 for the treatment of cancer. For example, teprotumumab targets the IGF-1 receptor (CD221) for the treatment of thyroid eye disease. For example, tesidorumab targets C5. For example, tetulomab targets CD37 for the treatment of cancer. For example, tezeperumab targets TSLP for the treatment of asthma and atopic dermatitis. For example, TGN1412 targets CD28 for the treatment of chronic lymphocytic leukemia and rheumatoid arthritis. For example, tibrizumab targets BAFF for the treatment of autoimmune diseases. For example, tildrakizumab targets IL-23 for the treatment of immunologically mediated inflammatory diseases. For example, tigatuzumab targets TRAIL-R2 for the treatment of cancer. For example, timigumab targets HER2 for the treatment of cancer. For example, timorumab targets AOC3. For example, tilagolumab targets TIGIT for the treatment of cancer. For example, tislerizumab targets PCDC1 and CD279 for the treatment of non-small cell lung cancer. For example, tisotumab vedotin targets coagulation factor III for the treatment of relapsed or refractory cervical cancer, and TNX-650 targets IL-13 for the treatment of Hodgkin lymphoma. For example, tocilizumab targets the IL-6 receptor for the treatment of rheumatoid arthritis. For example, tomzotuximab targets EGFR and HER1 for the treatment of cancer. For example, tralizumab targets CD154 (CD40L) for the treatment of rheumatoid arthritis and lupus nephritis. For example, tosatoxumab targets Staphylococcus aureus. For example, tositumomab targets CD20 for the treatment of follicular lymphoma. For example, tobetumab targets PDGFRA for the treatment of cancer. For example, tralokinumab targets IL-13 for the treatment of asthma and atopic dermatitis.For example, trastuzumab targets HER2 / neu for the treatment of breast cancer. For example, trastuzumab emtansine targets HER2 / neu for the treatment of breast cancer. For example, TRBS07 targets GD2 ganglioside for the treatment of melanoma. For example, tregalizumab targets CD4 for the treatment. For example, tremelimumab targets CTLA-4 for the treatment of non-small cell lung cancer, head and neck cancer, and urothelial carcinoma. For example, trevoglumab targets growth differentiation factor 8 for the treatment of orthopedic disuse and sarcopenia-related muscle atrophy. For example, tucotzumab cermoloukin targets EpCAM for the treatment of cancer. For example, tubirumab targets hepatitis B virus for the treatment of chronic hepatitis B. For example, ubrituximab targets MS4A1 for the treatment of multiple sclerosis and chronic lymphocytic leukemia. For example, urocuprumab targets CXCR4 (CD184) for the treatment of hematological malignancies. For example, urerumab targets 4-1BB (CD137) for the treatment of cancer. For example, urtoxazumab targets Escherichia coli for the treatment of diarrhea caused by E. coli. For example, ustekinumab targets IL-12 and IL-23 for the treatment of multiple sclerosis, psoriasis, and psoriatic arthritis. For example, utomirumab targets 4-1BB (CD137) for the treatment of diffuse large B-cell lymphoma. For example, badasutuximab talirin targets CD33 for the treatment of acute myeloid leukemia. For example, banalimab targets CD40. For example, polatuzumab vedotin targets STEAP1 for the treatment of cancer. For example, vanchikutumab targets the Frizzled receptor for cancer treatment. For example, vanucizumab targets angiopoietin 2 for cancer treatment. For example, bapariximab targets AOC3 (VAP-1). For example, valisakumab targets VEGF-A for the treatment of angiogenesis. For example, valrirumab targets CD27 for the treatment of solid tumors and hematological malignancies. For example, baterizumab targets ITGA2 (CD49b).For example, vedolizumab targets integrin α4β7 for the treatment of Crohn's disease and ulcerative colitis. For example, vertuzumab targets CD20 for the treatment of non-Hodgkin lymphoma. For example, bepalimomab targets AOC3 (VAP-1) for the treatment of inflammation. For example, besenkumab targets NRP1 for the treatment of solid malignancies. For example, bicilizumab targets CD3 for the treatment of Crohn's disease and ulcerative colitis. For example, bovali. Lizumab targets IL6R for the treatment of inflammatory autoimmune diseases. For example, boroximab targets integrin α5β1 for the treatment of solid tumors. For example, bonrelorizumab targets CD134 for the treatment of cancer. For example, voplaterimab targets CD278, also known as ICOS. For example, borsetuzumab mahodotin targets CD70 for the treatment of cancer. For example, botumumab targets the tumor antigen CTAA16.88 for the treatment of colorectal tumors. For example, bunakizumab targets interleukin-17 alpha. For example, zentuzumab targets IGF1 and IGF2. For example, XMAB-5574 targets CD19 for the treatment of diffuse large B-cell lymphoma. For example, saltumumab targets EGFR for the treatment of squamous cell carcinoma of the head and neck. For example, zanorimumab targets CD4 for the treatment of rheumatoid arthritis, psoriasis, and T-cell lymphoma. For example, zatuximab targets HER1 for the treatment of cancer. For example, xenoctuzumab targets ERBB3 and HER3 for the treatment of cancer. For example, diralimumab targets CD147 (basidine). For example, zolbetuximab targets CLDN18 for the treatment of cancer. For example, zolimomab aritox targets CD5 for the treatment of systemic lupus erythematosus and graft-versus-host disease.

[0194] In some embodiments, the antibody or antibody fragment may be human. Alternatively, the antibody or antibody fragment may be from a mouse. In some embodiments, the antibody or antibody fragment may be humanized.

[0195] In some embodiments, the antibody or antibody fragment may bind to a protein selected from Table 2. In some embodiments, the antibody or antibody fragment may bind to a protein encoded by IL2 (interleukin 2, ENSG00000109471). In some embodiments, the antibody or antibody fragment may bind to a histone complex. In some embodiments, the antibody or antibody fragment may bind to a protein encoded by kallikrein (KLK, ENSG00000167759). In some embodiments, the antibody or antibody fragment may bind to amyloid. In some embodiments, the antibody or antibody fragment may bind to a Notch receptor. In some embodiments, the antibody or antibody fragment may bind to a protein encoded by oxidized low-density receptor 1 (OLR1, ENSG00000173391).

[0196] 3. Signal transduction pathways The engineered platelets described herein may involve genetic modifications within the genetic components of the pathways for platelet adhesion, migration, and extravasation, or engineered platelets may be loaded with proteins, nucleic acids, or small molecule drugs. Engineered platelets may not respond to intrinsic stimuli that normally lead to clot formation, may not be recruited by other activated platelets, and may not be able to recruit and activate the patient's endogenous platelets upon activation.

[0197] Alternatively, deletions or modifications are introduced into genes that express thrombotic mediators such as HPS (biosynthesis of lysosomal organelle complex 3 subunit), which is essential for the release of ADP, serotonin, and ATP from dense granules, as well as genes that mediate platelet signaling, such as mitochondrial-encoded cytochrome C oxidase II (COX2), which generates inflammatory and pro-thrombotic mediators and is a target of aspirin. Alternatively, deletions or modifications are introduced into genes that express thrombotic mediators such as prothrombin (a major protein thrombotic inducer), the pro-angiogenic factors PDGF and EGF (elongation growth factor), and von Willebrand factor (collagen adapter protein).

[0198] While loss of thrombin and ADP signaling in combination has been observed to invalidate vascular occlusion, the ITAM receptor can still be activated (see Boulaftali et al. “Platelet ITAM signaling is critical for vascular integrity in inflammation”. JCI, 2013 and Cornelissen et al. “Roles and interactions among protease-activated receptors and P2ry12 in hemostasis and thrombosis”, PNAS. 2010, each of which is incorporated herein by reference). This study demonstrates that disruption of a critical endogenous platelet signaling pathway does not invalidate the ability of platelets to signal via the ITAM receptor, suggesting that the engineered CPR described herein is likely to function in a non-thrombotic platelet background.

[0199] For example, thrombin activates platelets through the cleavage of PARs (protease-activated receptors). Platelet signaling is also driven by protease-activated GPCRs, namely PAR1 and PAR4, which are cleaved by thrombin. The signaling is potent and acts to recruit platelets and promote positive feedback between platelets after platelet activation. The thrombin-cleaving sequences of PAR1 and PAR4 are well defined.

[0200] In some embodiments, the engineered platelets described herein may include at least one deletion or modification introduced or substituted into the domain of an endogenous platelet receptor, such as PAR4 (protease-activated receptor 4), a primary thrombin receptor; GP1b-1X-V (glycoprotein Ib complexed with glycoprotein IX), a primary anchor receptor; P2Y12 (purine receptor P2Y12), an ADP (adenosine diphosphate) receptor and target of clopidogrel inhibition; GPVI (glycoprotein deletion thein VI platelet), a collagen receptor; or a thromboxane receptor to prevent activation of the engineered platelets.

[0201] In some embodiments, manipulated platelets can synthesize proteins in response to activation signals. For example, Weyrich et al. showed that BCL-3 was specifically upregulated in activated platelets via a mechanical target of a rapamycin (mTOR)-dependent signaling mechanism (see Weyrich et al., "Signal-dependent translation of a regulatory protein, Bcl-3, in activated human platelets," PNAS, 1998, which is incorporated herein by reference in its entirety). Thus, gene knock-in to the BCL-3 locus or identification of the smallest 5'UTR region mediating activation-dependent translation enables regulation of synthetic gene expression in platelets. Therefore, the platelets described herein may have altered signaling pathways resulting in signal-inducible protein translation, for example, expressing toxic proteins once activated or triggering downstream events following recognition of target cells.

[0202] 4. Proteins related to autoimmunity In some embodiments, the manipulated platelet CPR described herein may include at least some of the proteins associated with autoimmunity. For example, the CPR may include myelin oligodendrocyte glycoprotein (MOG), glutamate decarboxylase 2 (GAD65), myelin-associated glycoprotein (MAG), peripheral myelin protein 22 (PMP22), thyroid peroxidase (TPO), voltage-opening potassium channel (VGKC), proteolipid protein (PLP), acetylcholine receptor (AChR), tribulus pseudokinase 2 (TRIB2), N-methyl-D-aspartate (NMDA) type glutamate receptor (GluR), glutamate decarboxylase 2 Carboxylase 2 (GAD2), Armadillo Repeat 9 (ARMC9), Cytochrome P450 Family 21 Subfamily A Member 2 (CYP21A2), Calcium-Sensing Receptor (CASR), Nuclear Autoantigenic Sperm Protein (NASP), Insulin, Thyroid-Stimulating Hormone Receptor (TSHR), Tyloperoxidase, Asioglycoprotein Receptor, Cytochrome P450 Family 2 Subfamily D Member 6 (CYP2D6), Lactoferrin (LF), Tissue Transglutaminase (TTG), H / K ATP-ase, Factor XIII (F8), Beta-2-glycoprotein I (Beta-2-GPI), Erythrocyte I / I, B2 integrin (ITGB2), Granulocyte colony-stimulating factor (G-CSF), Glycoprotein (GP) IIb / IIa, Collagen II (COLII), Fibrinogen (FBG) βα, Myeloperoxidase (MPO), Cardiac myosin (CYO), Proteinase 3 (PRTN3), Trichohyalin (TCHH), Bullous pemphigoid-related (BP), Glycoprotein 1 (GP1) It may contain at least a portion of proteins selected from the group consisting of laminin-332 (LM332), transglutaminase (TGM), type VII collagen (COLVII), P80 coilin (COIL), desmoglein I (DSG1), desmoglein III (DSG3), SRY-box 10 (SOX10), nuclear small molecular weight ribonucleoprotein U1 subunit (70SNRNP70), S antigen (SAG), and collagen α-3(IV) chain (α3(IV)NC1 collagen).For example, desmoglein 3-ITAM CPR may be used to target pemphigus vulgaris B cells. Alternatively, the engineered platelets described herein express an MHC class 1-ITAM chimeric platelet receptor or an MHC class 2-ITAM chimeric platelet receptor so that MHC class 1 or MHC class 2 can load peptides from the above list onto the surface of platelets that target autoimmune-mediated T cells for destruction or suppression by the release of anti-inflammatory cytokines such as TGF-β. Furthermore, RNA encoding transcription factors such as FOXP3 may be released to convert bound T cells into TRegs.

[0203] C. Universal platelets In some embodiments, the manipulated platelets described herein are less immunogenic than platelets produced in vivo (e.g., platelets from human donors). In vitro platelets can be made universal by deletion of the β2 microglobulin gene (see Feng et al. "Scalable Generation of Universal Platelets from Human Induced Pluripotent Stem Cells," Stem Cell Reports, 2014, which is incorporated herein by reference in its entirety). Even without this deletion, ABO-matched platelets are generally used in clinical practice without side effects. Human-derived type O platelets are not universal donors because they are contaminated with anti-A / B antibodies, but contamination is not a problem with in vitro platelets. Therefore, in certain embodiments, the inventions described herein can be used to mass-produce gene-edited platelets, which are also easily stored, transported, and do not require patient matching.

[0204] D. Cargo Platelets spontaneously absorb drugs and antibodies from the environment via endocytosis and open tubule systems (see Xu et al. "Doxorubicin-loaded platelets as a smart drug delivery system: An improved therapy for lymphoma." Scientific reports, 2017 and Verheul et al. "Platelets Take Up the Monoclonal Antibody Bevacizumab." Human Cancer Biology, 2007, which are incorporated herein by reference in their entirety). Platelets can be used to passively load and deliver genetically encoded therapeutic agents. For example, engineered platelets can be passively loaded with therapeutic agents via endocytosis and absorption. In fact, platelet α-granules contain protein effectors, and the loading of soluble proteins is carried out via simple signal peptides. The minimum target sequence for directing proteins to platelet-secreting α-granules has been previously defined (see Golli et al. "Evidence for a Granule Targeting Sequence within Platelet Factor 4," JBC, 2004, which is incorporated herein by reference in its entirety). In some embodiments, activation induces drug release in the manipulated platelet. The cargo may be soluble or membrane-bound.

[0205] Cargo can also be a contrast agent.

[0206] In some preferred embodiments, the cargo is not a drug naturally found within platelets; that is, the cargo is exogenous rather than endogenous with respect to platelets. Those skilled in the art will understand that the cargo may be exogenous with respect to platelets but endogenous with respect to the subject.

[0207] In some preferred embodiments, the cargo is not a drug naturally found within platelet α-granules. For example, the cargo may be a drug naturally found within platelets but not naturally found within α-granules.

[0208] In some embodiments, the cargo may be an agent found endogenously within platelets, or found in higher concentrations or amounts within platelets or within α-granules of platelets than in non-inventive platelets.

[0209] In some embodiments, the cargo includes an α-granule localization signal, which instructs the cargo to be taken up into α-granule vesicles of the manipulated platelet. For example, in some embodiments, a therapeutic agent or contrast agent includes or is complexed with an α-granule localization signal.

[0210] 1. Toxin In some embodiments, engineered platelets may be loaded with toxins that are coated from the immune system. Engineered platelets may be loaded with chemokines and / or selectins to mediate the transfer of drugs across the blood-brain barrier (BBB). Other embodiments of engineered platelets may have platelet secretory granules loaded with membranes and / or soluble proteins. In certain embodiments, the toxin may be encoded with an α-granule localization signal attached to direct its uptake into secretory granules, which will be released upon activation of the platelet receptor.

[0211] The expression of programmed cell death protein (PD-1) in platelets and the loading of engineered platelets with cyclophosphamide have been observed to function as potent anti-melanoma agents (see Zhang et al. "Engineering PD-1-Presenting Platelets for Cancer Immunotherapy." Nano Letters, 2018, which is incorporated herein by reference in its entirety). Specifically, megakaryocytes were engineered to express PD-1, and then cyclophosphamide was passively loaded into the resulting engineered platelets. Melanoma-targeting platelets are not synthesized receptors, but rather triggered in vivo by surgical wounds to the tumor, and tumor T reg Depletion and CD8 + This resulted in increased T cell-mediated cell death. Tumor volume was significantly smaller 20 days after the start of treatment in animals treated with both PD-1 and cyclophosphamide compared to animals treated with PD-1-expressing platelets or cyclophosphamide-loaded platelets.

[0212] 2. Nucleic acid and amino acid sequences In some embodiments, the cargo of the manipulated platelets of the present invention may be messenger RNA (mRNA). As used herein, the term “messenger RNA” (mRNA) refers to any polynucleotide that encodes a polypeptide of interest and can be translated to produce the encoded polypeptide of interest in vitro, in vivo, in situ, or ex vivo. Such mRNA molecules may have any structural components or features of those taught in International Publication No. WO2013 / 151666, which is incorporated herein in its entirety by reference.

[0213] c. CRISPR / Cas system In some embodiments, CRISPR / Cas gene editing systems can be used to modify the genome of megakaryocytes to generate the manipulated platelets described herein. Alternatively, CRISPR / Cas systems can be packaged in vesicles and released upon activation of platelets by antigens recognized by CRISPR. CRISPR / Cas systems are bacterial adaptive immune systems that utilize RNA-induced endonucleases to target specific sequences and degrade target nucleic acids. They are adapted for use in a variety of applications in the field of genome editing and / or transcriptional regulation. Any enzyme or orthologue known in the art or disclosed herein can be used in the methods herein for genome editing.

[0214] In certain embodiments, the CRISPR / Cas system may be a type II CRISPR / Cas9 system. Cas9 is an endonuclease that works with transactivated CRISPR RNA (tracrRNA) and CRISPR RNA (crRNA) to cleave double-stranded DNA. By ligating the 3' end of crRNA to the 5' end of tracrRNA with a linker loop, the two RNAs can be manipulated to form a single-molecule guide RNA. Jinek et al., Science, 337(6096):816-821 (2012), which is incorporated herein by reference in its entirety, demonstrates the usefulness of the CRISPR / Cas9 system for RNA-programmable genome editing, and international patent application WO2013 / 176772 provides numerous examples and applications of CRISPR / Cas endonuclease systems for site-directed gene editing, which are incorporated herein by reference in their entirety. Exemplary CRISPR / Cas9 systems include those derived from Streptococcus pyogenes, Streptococcus thermophilus, Neisseria meningitidis, Treponema denticola, Streptococcus aureas, and Francisella tularensis.

[0215] In certain embodiments, the CRISPR / Cas system may be a type V CRISPR / Cpf1 system. Cpf1, in contrast to the type II system, is a single RNA-induced endonuclease lacking tracrRNA. Cpf1 generates staggered DNA double-strand breaks with 4 or 5 nucleotide 5' overhangs. Zetsche et al. Cell. 2015 Oct 22;163(3):759-71, which provides examples of Cpf1 endonucleases usable in genome editing applications, is incorporated herein by reference in its entirety. Exemplary CRISPR / Cpf1 systems include those derived from Francisella tularensis, Acidaminococcus species, and Lachnospiraceae bacterium.

[0216] In certain embodiments, the specificity of CRISPR-mediated genome editing can be enhanced by using nickase variants of CRISPR / Cas endonucleases in which one or the other nuclease domain is inactivated. Nickase has been shown to promote HDR vs. NHEJ. HDR is directed from individual Cas nickases or using a pair of nickases adjacent to the target region.

[0217] In certain embodiments, a catalytically inactive CRISPR / Cas system can be used to bind to a target region (e.g., a gene encoding an antigen such as a receptor) and disrupt its function. Cas nucleases such as Cas9 and Cpf1 contain two nuclease domains. Mutations of key residues in the catalytic site create variants that bind only to the target site but do not result in cleavage.

[0218] In certain embodiments, the CRISPR / Cas system may include additional functional domains fused to the CRISPR / Cas endonuclease or enzyme. These functional domains may be involved in processes including, but not limited to, transcriptional activation, transcriptional repression, DNA methylation, histone modification, and / or chromatin remodeling. Such functional domains include, but are not limited to, transcriptional activation domains (e.g., VP64 or KRAB, SID or SID4X), transcriptional repressors, recombinases, transposases, histone remodelers, DNA methyltransferases, cryptochromes, photoinducible / controllable domains, or chemically inducible / controllable domains.

[0219] In certain embodiments, a CRISPR / Cas endonuclease or enzyme may be administered to cells or a patient as one or a combination of: one or more polypeptides, one or more mRNAs encoding polypeptides, or one or more DNAs encoding polypeptides.

[0220] d. Guide nucleic acids In certain embodiments, guide nucleic acids may be used to direct the activity of a relevant CRISPR / Cas enzyme to a specific target sequence within the target nucleic acid. Guide nucleic acids provide target specificity to the guide nucleic acid and CRISPR / Cas complex by binding to the CRISPR / Cas enzyme, and thus guide nucleic acids can direct the activity of the CRISPR / Cas enzyme.

[0221] In one embodiment, the guide nucleic acid may be an RNA molecule. In one embodiment, the guide RNA may be a single-molecule guide RNA. In one embodiment, the guide RNA may be chemically modified. In certain embodiments, two or more guide RNAs may be provided to mediate multiple CRISPR / Cas-mediated activities at different sites within the genome.

[0222] 3. Small molecule drugs In some embodiments, the cargo within the vesicles of the manipulated platelets described herein includes (-)-epigallocatechin 3-gallate, (-)-fencerin, (+)-calanolide A, (R)-folitixolin, (R)-mequitazine, (S)-pantoprazole sodium, [11C]DASB, [11C]-laclopride, [18F]FDG, [18F]HX4, 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea, 1,2-decanediol, 11,11-di-duteroethyllinoleic acid, 11C-PBR-28, and 123I-io Methopane, 124I-CLR-1404, 131I-MIBG, 131-iodine, 13-cis-retinoic acid, 13C-labeled metacetin, 13N-ammonia, 1400W94, 17-beta-estradiol, 17-alpha-hydroxyprogesterone caproate, 17-beta-estradiol, 17-beta-estradiol valerate, 17-hydroxysteroid dehydrogenase inhibitor, 18F-EF5, 18F-FDG, 2L polyethylene glycol, 25-dihydroxy-vitamin D3, 25-OH vitamin D, 2-chloroprocyan 2-Deoxyglucose, 2-Hydroxypropyl-Beta-Cyclodextrin, 2MD, 2-Methoxyestradiol, 4-Aminopyridine, 4-Aminosalicylic Acid, 4-FEC, 4-Hydroxytamoxifen, 5-Aminolevulinic Acid, 5-Aminosalicylic Acid, 5-Arachitaine, 5-Fluorouracil (5-FU), 5-Hydroxytryptophan, 5-Methoxypsoralen, 6-Mercaptopurine, 6-Thiogunine, 9-Aminocamptothecin, 9-Aminofucine, 9-Nitrocamptothecin, Abacavir, Abafungin, Abame Tapil, avediterol, avexinostat, abiraterone, ABT-072, ABT-751, akadesin, acalabrutinib, acamprosate, acamprosate calcium, acarbose, acevirstat, acebutrol, acexidine, aceclofenac, acenulamic acid, acenocoumarol, acetadothe, acetaminophen, bicarbonate without acetic acid, acetazolamide, acetic acid, acetylcholine, acetylcysteine, acetyl-L-carnitine, acetyl-L-carnitine hydrochloride, acetyl-L-leucine, acetylsalicylic acid,Acetyl salicylic acid, acetylsalicylic acid (ASA), acetylsalicylic acid ricinate, acyclovir, acipimox, acitretin, acralubicin, acridinium, acridinium bromide, acorbifen, acorafloxacin, acotiamide hydrochloride, ACP-104, acribastine, ACT-01, ACT-280778, actinomycin D, acumapimod, acillin, adapalene, ADC-3680, Adela XR, adefovir dipivoxil, ademethionine, adenosine, azinazolam, adipiplon, adme Glibant, Adzelesin, Adramycin, Adrenaline, Adrenaline, Adriamycin, Advea, Advil, AE-941, Afasifenacin fumarate, Afatinib, Afegostat, Apheretecan, Afimoxifen, Aflibercept, Aftovetin, Afresertib, Aganepagisopropyl, Agatrimod, Agaveinulin, Agomelatine, Alphagan, Ajumarin, Aladrian, Aragebrium chloride, Alanyl glutamine dipeptide, Albaconazole, Albendazole, Albiglutide, Albithiazolium Romido, Albumin, Albuterol, Albuterol Sulfate, Albuterpenoid, Alkaftazine, Alsipotriol / Betamethasone, Aldesleukin, Aldoxyrubicin, Alectinib, Allegritazar, Alemtuzumab, Alendronate, Alendronate Sodium, Alendronate, Alexell, Alive, Alfacalcidol, Alfentanil, Alfuzosin, Algerdrate / Magnesium Oxide, Alimta, Alicertib, Aliskiren, Alitretinoin, Alitretinoin, Alizaprid, Allantoin, Alisartan Isoproxil, allopregnanolone, allopurinol, all-trans retinoic acid, almorexant, almotriptan, alodane, agomelatine benzoate, alosetron, alovudine, alpelisib, alpha-lipoic acid, alpha-tocopherol, alpha-1 antitrypsin, alpha-cyclodextrin, alpha-glucosidase inhibitor, alpha-interferon, alpha-lipoic acid, alpha-tocopherol, alpha-tocopherol acetate, alpha-tricosanthin, alprazolam, alprostadil, alprostadil alfadex, ALS-08,Artinicline, Altropane, Aluminum MgS, Aluminum Hydroxide, Albespimycin Hydrochloride, Albimopan, Arbocidib, Amantadine, Amantadine Hydrochloride, Ambrisentan, Ambroxol, Ambroxol Hydrochloride, AMD-070, Amdoxovir, Amervant, Amenamevir, Ametop, Amphetamine, Amibegron, Amifampridine Phosphate, Amifostine, Amikasic Acid, Amyloride, Amyloride Hydrochloride, Amino Acids, Aminocaproic Acid, Aminoglutethimide, Aminoguanidine, Aminolevulin Acids, aminolevulinic acid hydrochloride, aminophylline, aminopterin, amiodarone, amiprilose, amicelimod, amisulfide, amitifazine hydrochloride, amitriptyline, amitriptyline hydrochloride, amlexanox, amlodipine, amlodipine besylate, amlodipine maleate, amlodipine besylate, amlodipine camusylate, amlodipine maleate, ammonium lactate, amnion, amodiaquine, amonafide dihydrochloride, amonafide L-malate, amorolphin, amoxapine, amoxicillin, amoxicillin Labranate, Amoxicillin MR, Amoxicillin / Clavulanate, Amoxicillin-Clavulanate, Amoxicillin, Amphetamine, Amphetamine Aspartate, Amphetamine Sulfate, Amphotericin, Amphotericin B, Ampicillin, Ampicillin Sodium, Ampicillin / Flucloxacillin, Amprenavir, Amrubicin, Amsacrin, Amcillarotene, AN-2898, AN-9, Anacetrapib, Anagliptin, Anagrelide, Anamorelin, Anastrozole, Anativant, Anclivirock, Anclod, An Drogen, Androxy, Anecoltabe, Angiotensin-Converting Enzyme Inhibitors, Angiotensin I, Angiotensin II, Angiozymes, Anidurafungin, Aniracetam, Anamycin, Antazolin, Anthocyanins, Anthracrine, Antiemetics, Antihistamines, Antilymphocyte Globulins, Antitumor Drug A-10, Antitumor Drug A10-I, Antitumor Drug AS2-1, Antioxidant Vitamins, Antipsychotics, Antiretroviral Drugs, Antithymocyte Globulins, Antithymocyte Globulins, Apabet, Apadenoson, Apadiquon, Aperin, Apheresis, ApirimodApimostinel, Apitricib, Apicaban, Apraviroc, Aprindre, Pomorphine, Apovir, Aplatastat, Apremilast, Aprepitant, Applicitabine, Appricoxib, Aprotinin, AR-623, Ara-C, Arachidonic acid, Arachitin, Aralast, Aramcol, Aracettaconazole, Albaclofen, Albaclofenplacarbil, Arbekacin sulfate, Arbutin, Arc-100, Alformoterol, Argatroban, Argatroban, Arginine, Arginine vasopressin, ARH-1, Alha Lophenate, alimocromol, aripiprazole, almodafinil, alogliptin, arsenic trioxide, artephenomer mesylate, artemether, artemether-lumefantrine combination, artemisinin, artemizone, artemotil, artenimol, arterolan, arterolane maleate, artemiartesunate, artesunate + mefloquine, artesunate-amodiaquine, articaine, articaine hydrochloride, arundic acid, alzoxifen, asapipant, ASCJ-9, ascorbic acid, ascorbic acid, asenapine, Asimadrin, ASM-024, asoprisnil, aspirin, astaxanthin, astodrimer, asunaprevir, AT-101, ataciguat, atagabalin, atallen, atamestan, atazanavir, atazanavir sulfate, atazanavir / ritonavir, atesegatran fexenetil, atervia, atenatib, atenolol, atevirdin, ATHX-105, atiprimod, atiratecan, atiban, atomoxetine, atopaxer, atorvastatin, atobakon, atracurium, atracurium besylate, atrasentan, atre Iton, Atripra, Atropine Auranofin, Auriclozen, AVAC, Abacopan, Abagasestat, Avanafil, Awacimib, Abatrombopag, AVE-0657, AVE-2268, Avibactam sodium, Avil, Avobenzone, Avoralstat, Avosentan, AWD-12-281, Axeroplan, Axilon, Axitinib, Axomadol, Azacitidine, Azathioprine, AZD-1775, AZD-4547, AZD-9668, Azedra, Azelaic acid, Azelastine, Azelastine hydrochloride, Azerilagon,Azelnidipine, azidothymidine, azilsartan, azithromycin medoxomyl potassium, azimilide, azithromycin, azithromycin dihydrate, azosemide, aztreonam, aztreonamidine, bacitracin, baclofen, bafetinib, baicalin, paraglitazone, valacatib, valsalazide, bambuterol, banoxantrone, valacetib, bardoxolone methyl, valcitinib, barnidipine, basiliximab, basimugulanto, basumisanil, batabulin, batefenterol succinate, babisant, bazedox Cyfen, BCG vaccine, BCNU, becatecarin, beclabuvir, beclomethasone, beclomethasone dipropionic acid, beclomethasone, beclomethasone dipropionic acid, becocalcidiol, beconase, bedaquiline, vedradrine, vedrocone, bellinostat, belladonna, vernacasan, beroranib, berotecan, bempedonic acid, benazepril, bendamustine, bendroflumethiazide, bene, benfotiamine, benidipine, bensazide, bentamapimod, benzalkonium, benzalkonium chloride, benzathine penicillin, benzbroma Ron, benznidazole, benzocaine, benzodiazepine, benzophenone-3, benzoyl peroxide, benztropin, benzydamine hydrochloride, benzyl alcohol, benzylpenicillin, benzylpiperazine, bepanthol, bepotastine, belatant, beraprost sodium, berberine, berbicin, besifloxacin, besifovir, betaerythropoietin, beta-1,3 / 1,6-glucan, betablocker, betablocker, betacarotene, betacryptoxanthin, betadine, betafectin, betahistine, beta In, anhydrous betaine, beta-lactamase inhibitor, betamark, betamethasone, betamethasone dipropionate, betamethasone mousse, betamethasone valerate, betamethasone dipropionate, tricalcium phosphate bone substitute, betaxolol, betaxolol hydrochloride, betanethyl, betanethyl chloride, betrixaban, betulinic acid, bevacizumab, bebenoplan, bevirimat, bexagliflozin, besarotene, bezafibrate, BF-derm1, BGP-15, BI-54903, biapenem, bicalutamide, bififadin, bifeprnox,Bifidobacterium, Bifidobacterium bifidum, Bifidobacterium infantis 35624, Bifonazole, Biguanide, BIIB-021, Bilastine, BILR-355-BS, Bimatoprost, Bimocromol, Bimosiamors, Vindalit, Binimethinib, Binode, Noson, Bio-25, Biotin, Biperiden, Bifentin, Virabrecib Dihydrate, Bilicodal, Birinapant, Bisacodyl, Biscarcitol Potassium, Bismuth, Bismuth Citrate, Bismuth Citrate Potassium, Bismuth Sodium Tartrate, Bismuth Subcitrate, Bismuth Subsalicylate, Bisoprolol, Bisoprolol Fumarate, Bisphosphate, Bitpertine, Bixalomer, Bleomycin, Bleomycin Sulfate, Blonanserin, BMP-7, BNC-105P, Boceprevir, Boric Acid, Boron Anticancer Drug, Bortezomib, Bocenta Bosutinib, bladaniclinn, bradykinin, bramitob, branched-chain amino acids, brekanavir, brexiprazole, bricanyl, bricasol, brimonidine, brimonidine tartrate, brinaves, brinzolamide, brivanib alaninate, brivalacetam, brivudine, brolucizumab, bromfenac, bromfenac sodium, bromhexine, bromocriptine, brompheniramine, bronopol, brostarisin, brotizolam, briostatin-1, BTI-320, BTL-TML-HSV, BTS-67582, bucindol, b Desonide, budesonide / formoterol, budesonin, budiodaron, bumetanide, bunazosin, buparlisib, bupivacaine, bupivacaine hydrochloride, bupivacaine with fentanyl, bupivacaine-clonidine, buprenorphine, buprenorphine hemiadipate hydrochloride, buprenorphine hydrochloride, buprenorphine / naloxone, bupropion, bupropion hydrochloride, bupropion SR, brapitant, blixafor, busererin, busererin acetate, buspirone, buspirone hydrochloride, busulfan, busulfex, butarvital, bu Tenafine, Butoconazole, Butoconazole Nitrate, Butorphanol, Butorphanol Tartrate, C5a, Cabaseryl, Cabazitaxel, Cabergoline, Cabotegravir, Cabozantinib S-Malate, Cassit D3, Cadazolide, CAF Regimen, Caffeic Acid, Caffeine, Caffeine Citrate, Caffeinol, CalciTur D3 Forte, Calcipotriol / Beta-Methasone, Calcitriol, Calcium, Calcium Acetate, Calcium Ascorbate, Calcium Carbonate, Calcium Chloride, Calcium Chloride Dihydrate, Calcium Citrate,Calcium dobesylate, calcium fluoride, calcium folate, calcium glucarate, calcium gluconate, calcium hydrogen phosphate, calcium L-aspartate, calcium levofolate, calcium phosphate, polycarbophil calcium, sodium calcium phosphate, calcium supplements, calcium and vitamin D, calcium leucovorin, caldalet, calfactant, camisinal, camobucol, camptothecin, canagliflozin, candesartan, candesartan cilexetil, canative, camphosphamide, Cangrelol, cannabidiol (CBD), cannabidivarin, cantharidin, capadenoson, capecitabine, camatinib, capolac, caplavillin, capros, capsaicin, captopril, carbamazepine, carbenoxolone, carbetimer, carbetocin, carbidopa, carbocysteine, carbocysteine, carbogen, carbon[14C]oxaliplatin, carbon dioxide, carbon monoxide, carbon dioxide, carboplatin, sodium carboxymethylcellulose, cardidopa, cardinutrient, carfilzomib, cargrumic acid, caripolide, kali Plazin, Carisbamate, Carisoprodol, Carmegliptin, Carmoterol, Carmustine, Carnitine, Carotegras Methyl, Carteolol, Carteolol Hydrochloride, Carvedilol, Carvedilol Phosphate, CASAD, Casein, Casopitant, Caspofungin, Catetine, CBT-1, CCPI, Cebranopadol, Sediranib, Cefaclor, Cefadroxil, Cephalexin, Cefazolin, Cefazolin Sodium, Cefdinir, Cefditoren Pivoxil, Cefepime, Cefirabancin, Cefixime, Cefmetazole, Cef Fuperazone, cefotaxime, cefotetan, cefoxitin, cefozopran, cefpirome, cefpodoxime, cefprodil, cephthaloline, cephthalolinefosamil, ceftazidime, ceftibuten, cergocivir, celiprolol, ceribaron, senestin, senicliviroc, cenovamate, sensabudine, centanafadine, centanafadine, centramforte, centanasilver, cephalexin, cephalosporin, cerarifimod, ceramide, ceritinib, cerium nitrate, cerivastatin, serapyridine, certoparin, cetaphil, cethromycin,Cetilistat, cetirizine, cetylpyridinium chloride, cevimeline, chenodeoxycholic acid, CHF-1535, CHF-5992 chitosan, chitosan-thiomer, chlorambucil, chloranphenicol, chlordiazepoxide, chlorhexidine, chlorhexidine, chlorhexidine digluconate, chlorhexidine gluconate, chlorhexidine alcohol, chlorhydrate, chlormadinone acetate, chloroprocaine, chloroquine, chloroquine diphosphate, chloroquine phosphate, chlorpheniramine, chlorpheniramine maleate, Lorproguanil, chlorpromazine, chlorthalidone, chlorzoxazone, cholecalciferol, cholecystokinin-8, cholesterol absorption inhibitors, cholestyramine, cholic acid, choline, choline alphoserate, cholinediepalestat, choline fenofibrate, magnesium trisalicylate choline, chondroitin, chondroitin sulfate, CHP-HER2, cobalt chromium, chromium picolinate, CHS-131, CHVP-interferon, ciclesonide, cicletanin, cyclopirox, cyclosporine olamine, cyclosporine, Cyclosporine A, cidofovir, silancetron, cilastatin, cilazapril, cilnidipine, siromilast, cilostazol, silprevir, cimetidine, simicoxib, cinacalcet, cinaciguat, sindunistat hydrochloride maleate, cinnamaldehyde sodium hyaluronate, sinitaprid, sinitaprid tartrate cinnamaldehyde, cinnarizine, sipalgamin, ciprofiflate, ciprofloxacin, ciprofloxacin hydrochloride, silaparantag, cisapride, cisatracrium, cisatracrium besylate, cisplatin, cisplat Tin liposomes, cisplatinum, citalopram, citalopram hydrobromide, Citanest, citicoline, citric acid, fentanyl citrate, citric acid, citric acid monohydrate, citrulline, CK-2017357, cladribine, Clarinex, clarithromycin, clavulanate, clavulanate potassium, clavulanic acid, clazosentan, clevoprid, clemastine, clemastine fumarate, clenbuterol, clebidipine, clevudine, clindamycin, clindamycin phosphate, clindamycin / benzoyl peroxide,Clinisole, Cryoquinol, Clobazam, Clobetasol, Clobetasol propionate, Clobex, Clodronate, Clofarabine, Clofazimine, Clomethiazole edicilate, Clomifene, Clomifene, Clomifene citrate, Clomifene, Clomifene citrate, Clomipramine, Clonazepam, Clonidine, Clonidine hydrochloride, Clopidogrel, Clopidogrel bisulfate, Clopidogrel napadisylate, Chlorat D, Clorazepate, Clostridium butyrachum MIYAIRI 588, clotrimazole, cloxacillin, clozapine, coamoxiclaf, cobalamin, cobamamide, cobicistat, cobimetinib, cobiprostone, cocaine, codeine, codeine phosphate, coenzyme Q10, coeruleus, cogentin, cogentus, colchicine, colsine, cholecalciferol, cholecalciferol D3, collenuramide, colseveram, cholestylan, cholestipol, colfoseryl palmitate, colestimetate, sodium colimethicine, colistin, colistin sulfate, colistineb, colloidal bismuth Colloidal bismuth tartrate, Coroxyl, Colracetam, Compretastatin, Comp-01, Comp-02, Comp-03, Comp-04, Conivaptan, Conivaptan hydrochloride, Conjugated estrogen, Conjugated estrogen, Sustained-release carvedilol, Copanlisib, Copper, Copper histidine, CoQ10, Cortexolone 17α-propionic acid, Corticosteroid, Cortisol, Cortivazole, Cocitecan, Cosmopha, Cotrimoxazole, Cotrimoxazole, Cotrimoxazole, Cotrimoxazole, COX inhibitor, CPI-613, CRAd 3 / 5-Delta, Creatine, Creatine Ethyl Ester, Creatine Monohydrate, Crenolanib, Chrysabolol, Crizotinib, CRM-197, Clobenetin, Cloferemer, Cromoglycate, Cromoglycinic Acid, Cromolyn Sodium, Cresitaloid, C-Tb, CTO, CUDC-305, Curcuma eruginosa, Curcoumarin, Curcuminoid, Cardran Sulfate, Katamecin Dihydrochloride, CX-516, Cyanocobalamin, Cyclodine, Cyclodine Lactate, Cyclobenzapurine, Cyclobenzapurine Hydrochloride, Cyclodextrin, Cyclodextrin-bound Diclofenac,Cyclogest, cyclopentolate, cyclophosphamide, cyclophosphamide, cyclophosphamide monohydrate, cyclophosphan, cycloserine, cyclosporine, cyclosporine, cyclosporine A, cyclosporine microemulsion, cyclophosphamide, cyproheptadine, cyproterone acetate, cyproterone, cyproterone, cyproterone acetate, cysteamine, cysteamine hydrochloride, cysteine, cysteine ​​hydrochloride monohydrate, cytarabine, cytarabine arabinoside, cytarabine-asparagine conjugate, cytophosphan, cytosine-arabinoside Vinoside, Cytosine Arabinoside, Cytoxan, Cytozar, D3 Vitamin, DA-9601, Dabigatran Etexylate, Dabrafenib, Dacarbazine, Daclatasvir, Daclizumab, Dacomitinib, Dactinomycin, Dactricib, Daglutril, Daidzein, Dalbavancin, Dalcetrapib, Dalphopristin, Dalteparin Sodium, D-Amphetamine, Danazol, Danilixin, Danoprevir, Dantrolen, Dantrolen Sodium, Danacertib, Dapaconazole, Dapagliflozin, Dapagliflozin Propanedio Lu, dapanstril, dapivine, dapolinad, dapoxetine, dapsone, dapsone gel, dalapradib, dalifenacin, darinaparcin, darolutamide, darotropicin bromide daunorubicin, darunavir, darunavir / ritonavir, darzentan, dasabuvir, dasatinib, dasotraline, daunorubicin, daunorubicin hydrochloride, daunorubicin, D-cycloserine, dDAVP, DDP, DE-104, DE-110, DE-112, Deanxit, Debio-1450, Debio-1452, Decadron, decarbazine, desernotinib, Decitabine, Decoglucant, Decuprate, Defactinib, Deferasirox, Deferipron, Deferitasol, Deferoxamine, Deferoxamine Mesylate, Deflazacort, Dehydroepiandrosterone, Delafloxacin, Delamanid, Delanzomib, Delapril, Delapril Hydrochloride, Delavirdine, Delazin, Deleobvir, Deligopalin Sodium, Delorazepam, Delta-8-THC, Delta-9-Tetrahydrocannabinol, Denagliptin, Denufosol Tetrasodium, Depacon, Depade, Depomedroxyprogesterone,Depomedroxyprogesterone acetate, depomethylprednisolone, depotestosterone, DER-45-EV, derenophylline, delsalazine sodium, desferrioxamine, desflurane, desipramine, desloratadine, desmopressin, desmopressin acetate, de, Sogestrel, desonide, desotefumarate, deoxymethasone, desvenlafaxine, detox B adjuvant, deutetrabenazine, dexamethasone, dexamethasone acetate, dexamethasone cypecylate, dexamethasone diphosphate, dexamethasone phosphate, dexamethasone sodium phosphate, dexamphetamine, dexanabinol, dexachlorpheniramine, doxiderin, dexerbucitabine, dexifenflamin, dexibuprofen, doxide, deximethene mucate, dexketoprofen, dexketoprofen trometamol, dexlansoprazole, dexlipotam, dexroxyglumide, dexmecamylamine, dexmedetomidine, dexmethylphen Date, Dexnigludipine, Dexpanthenol, Dexramipexol, Dexrazoxane, Dexrazoxane Hydrochloride, Dextophysopam, Dextran, Dextroamphetamine, Dextroamphetamine Saccharate, Dextroamphetamine Sulfate, Dextromethorphan, Dextromethorphan Hydrobromide, Dextropropoxifen, Dextropropoxifen Hydrochloride, Dextrose, Dexverapamil, Dezosine, DHEA, Diaselein, Diacetylmorphine, Diamel, Diamine Dichloride Platinum, Diamorphine, Diamorphine Hydrochloride, Dihydrogalactitol, Dianiclean, Diao Xin Xue Kang), diazemul, diazepam, diazepam autoinjector, diazoxide, diazoxidecholine, dibasic sodium dihydrate phosphate, dibekacin, diclofenac, diclofenac diethylamine, diclofenac potassium, diclofenac sodium, dicloxacillin, didanosine, dienogest, diethylcarbamazine, diethylnorspermine, diethylpropion, diethylstilbestrol, diflomotecan, diflunisal, difluprednate, digoxin, digoxin, dihematoporphyrin, dihomo-γ-linolenic acid, dihydrazine, dihydroartemisinin, dihydroartemisinin-piperaquine, dihydrocodeine, dihydroergotamine, dihydroergotamine mesylate, dihydroxyvitamin D3,Diiodotyropropionic acid and its analogues, diphenhydramine, dilaudide, dilmapimod, diltiazem, diltiazem hydrochloride, dimenhydrinate, dimenhydrinate, dimesuna, dimethindene, dimethindene maleate, dimethyl fumarate, dimethyl fumarate, dimiracetam, inoprostone, diosmin, diphencypron, diphenhydramine, diphenylcyclopropenone, dipyrone, dipragurant-IR, dipyradimol, dipyridamole, dipyrone, diquafosol sodium, diquafosoltetrasodium, disopyramide Dispase II, disfenton sodium, disulfiram, dislanol, dithiocarb sodium, DLBS-1033, DLBS-1425, D-methadone, DNE3, dobutamine, docetaxel, dosipalstat, doxorubicin, doxorubicin ethyl ester, doxorubicin acid [DHA], doxorubicin acid monoglyceride, docosanol, doxorubicin, doxorubicin sodium, dofetilide, drasetron, dorastatin-10, dorogesic, dolutegravir, domperidone, donepezil, donepezil hydrochloride, donu, Dopamine, dopexamine, dramapimod, doravirine, doripenem, dorzolamide, dorzolamide hydrochloride, dorzolamide hydrochloride + timolol maleate, dotiepin, dovitinib, doxapram, doxazosin, doxorubicin mesylate, doxepin, doxepin hydrochloride, doxelciferol, doxifluridine, doxil, doxophilin, doxorubicin, doxorubicin etalforatide, doxorubicin HCl liposome, doxorubicin hydrochloride, doxorubicin hydrochloride liposome, doxycycline, doxycycline hydrochloride, doxyl Min, doxylamine succinate, D-penicillamine, DPP-IV inhibitor, DPS-102, draflazin, dorinabant, dronabinol, dronedarone, droperidol, dropyropidine, drospirenone, dorotaverin, droxidopa, D-tagatose, D-trans fentanyl, Duac, dual-release hydrocortisone, dulaglutide, dalcolax, duloxetine, dulacaine, dulamorph, duloran, dutasteride, dutogliptin, duvelisib, duvoglustat, D-xylose, dydrogesterone,Dihydroprogesterone, DZ-1, E. Corinissle, E-7016, E-7820, Ebastine, Ebselen, EC-17, Ecabet, Ecabet sodium, Echinacea, Econazole nitrate, Ecopipam, Ecosprine, Eclaprost, Eculal, Edaglitazone, Edaravone, Calcium disodium edetate, Ediboxetine, Ednerpic maleate, Edtecarin, EES0000645 / A, Efaproxial, Efaturtazone, Efavirenz, Efinaconazole, Efronitine, Efonidipine hydrochloride, EGb-761, EGCG, Eicosapentaenoic acid, Eicosapentanoic acid, Elastolant, Elacridal, Elacitarabine, Elafibranol, E Lagolix, ellamipretide, elbasvir, elvion, eldecalcitol, eleclazine, elethcromol sodium, eletriptan, eliglustat tartrate, erinogrel, eriprodil, ELND-005, elobixibat, elocalcitol, elomet, erosalic, erusamitrusine, eltoprazine, eltrombopag, elbrixin, erbucitabine dihydrochloride, elvitegravir, ervorin, erbucitabine, elzasonan, emdogain, emedastine, emepimut-S, emicellphone, emibilin, emmixstat, empagliflozin, emricasan, emtricitabine, enalapril, enalapril maleate, enasidenib. Encaret sulfate, enclomifene citrate, enclomifene, encorafenib, endocannabinoid palmitoylethanolamide, endonase, endotoxin, cyclophosphate, enecazin, enflurane, enfuvirtide, enyluracil, ENMD-2076, enobotherm, enocitabine, enoxaparin sodium, enoximon, entacapone, entecavir, entecavir maleate, enteric-coated mycopheno Sodium lat, enteric-coated tegafur-uracil, enteraggregating Escherichia coli, entinostat, Entonox, enzalutamide, Enzasta Epadel, Epalestat, Epadel, Epalestat, Eperisone, Eperisone hydrochloride, Epetirimod, Epetrabolol, Ephedrine, Epiceram, Epidoxirubicin, Epidoxorubicin, Epidural / paraspinal analgesia, Epigallocatechin gallate, Epigallocatechin-3-gallate,Epigallocatechin gallate, epigallocatechin, epinastine, epinastine hydrochloride, epinephrine, epirubicin, epirubicin hydrochloride, episalvan, eplerenone, eprivanserin, epoprostenol, epicajutsu, eprodicate, eprosartan, eprotilome, epsilon-aminocaproic acid, eptastigmine, eptifibatide, equine antithymocyte immunoglobulin, Equisetum arvense, ellabacycline, erdostein, Eremostachys laciniata, ergocalciferol, ergotamine, eribaxaban, eribulin mesylate, Elitex, eritoran, erlotinib, ertapenem, erteberel, ertugliflozin, erythromycin, erythromycin lactobionic acid, erythropoietin, erythropoietin beta, ESAT-6CFP10, esaxerenone, E. coli endotoxin, escitalopram, esflurbiprofen, es Caris, Esketamine, Esketamine Hydrochloride, Eslicarbazestepin Acetate, Esmeron, Esmirtazapine, Esmolol, Esmolol Hydrochloride, Esomeprazole, Esoxibutynin, Eslevocetin, Estazolam, Estrol, Estradiol, Estradiol Acetate, Estradiol Cypionate, Estradiol Valerate, Estradiol / Norethindrone Acetate, Estramusti Estramustine phosphate, sodium estramustine phosphate, estrates, estriol, estriol E3, estrodiol, estrogen, estrogen, ezopiclone, ethacrinic acid, etalosib, etalfa, etanercept biosimilar, ethacolate, ethacric acid, ethambutol, ethambutol hydrochloride, ethanol, ethinylestradiol, ethinylestradiol / levonorgestrel, ethinylestradiol, ethiodidine oil, etosquimide, ethyl chloride, ethyl eicosapentaenoate, ethylcalcium hydrogen fumarate, ethylmagnesium hydrogen fumarate, ethylzinc hydrogen fumarate, ethylenediaminetetraacetate, ethylhexyl triazone, ethinylcytidine, etidronic acid, etilephrine, etodrug, etomidate, etomidate tripro, etomoxyl, etonogestrel,Etonox, Etoposide, Etoposide Phosphate, Etricoxib, Etravirine, EV-06, Evatetrapib, Evatanepag, Everolimus, Eviprostat, Evophosphamide, Evogliptin, Exatecan, Exemestane, Exenatide, Exeporfinium Chloride, Exisrind, Estradiol, Ezetimibe, Ezetimibe / Atorvastatin, F-0434, F0-M0, F0-M1700, F160-M0, F160-M1000, F160-M1700, F80-M1000, F80-M1700, FA, Fam Hydrochloride Cyclovir, famciclovir, famciclovir, famitinib L-malate, famotidine, fanpridin, fandi, phalampeter, farglitazar, faropenem, faropenem medoxomil, faciglifam hemihydrate, facitibant chloride, fasudil, favipiravir, FBG-18, FBP peptide, FE

[50] C, FE

[75] C, Fe-58, febuxostat, fedbapagon, phedratinib, felbamate, felbinac, felodipine, phenatonil, phenobam, fenofibrate, phenold Pam, fenoterol, fenoterol prednisone, fenretinide, fentanyl, fentanyl citrate, fermadic acid, ferric carboxymaltose, iron citrate, iron maltol, iron pyrophosphate, ferlipel-3, ferroquine, iron fumarate, iron sulfate, fermoxtran-10, fermoxyitol, FeSO4, fesoterodine fumarate, febipiprant, fexinidazole, fexofenadine, fivoflapon, fibrinogen, fidaxomicin, filanesib, filgotinib, filgrastim, filibuvir, firo Lexant, Fimaporfin, Fimasartan, Finafloxacin, Finafloxacin hydrochloride, Finasteride, Filenone, Fingolimod, Fioliset, Fipamezole, Fish oil (eicosapentaenoic acid [EPA] + docosahexaenoic acid [DHA]), Physiogel, Fispemifene, Frazil, Flavan-3-ol, Flavanone, Flavoxate, Flecainide, Flibanserin, Flomoxef, Flomoxef sodium, Flopristin, Florbenazine, Florbetapir, Flortausipil F18, Floric acid, Flobagatran,Floroxlysine, flucyclatide F18, flucloxacillin, fluconazole, flucytosine, fludarabine, fludeoxyglucose, fludeoxyglucose F18, fludrocortisone, flumazenil, flunarizine, flunisolide, fluocinolone acetonide, fluocinonide, fluorescein, fluorometholone, Fluorometholone acetate, fluoropyrimidine, fluoroquinolone, fluorouracil, fluoxetine, fluoxetine hydrochloride, flupentixol, fluphenazine, flupirtin, flurbiprofen, flurbiprofen axetil, flurbiprofen sodium, flutamide, fluticasone, fluticasone furoate, fluticasone propionic acid, flutrimazole, fluvastatin, fluvoxamine, FM-VP4, folacin, folic acid, folic acid / iron, FOLFIRI, FOLFOX4, FOLFOXIRI, folic acid, folic acid, folic acid Follicoxolin calcium, follitropin beta, fonadelpar, fondaparinax sodium, folazyl, foretinib, formoterol, formoterol fumarate, folodestine, folopafant, fosalvudine tidoxyl, fosanprenavir, fosanprenavir calcium, fosaprepitant, fosbretabrin, fosbretabrin disodium, foscarnet, foscarnet sodium, phosphagrocoat, fosdevin, fosfluridine tidoxyl, phosfomycin, phosfomycin trometamol Phosfructose, Hosinopril, Fosmidomycin, Fosphenytoin, Phospropofol, Fostamatinib, Fostemsavir Tromethamine, Fostimon, Fostrap, Hotemustine, Hodivudine Tidoxyl, Frezeresta, Fresvin, Flovatriptan, Fructose, Fructose-1,6-Diphosphate, Fluquintinib, Flusemide, Fucoidan, Fulvestrant, Fumarate, Funapide, Flaprevir, Furazolidone, Furosemide, Fusidetate Sodium, Fusidic Acid, Fuzenfayu, Gabapentin, Gabapentin Enacarbil, Gaboxadol, Gadopentate, Gadobutrol, Gadophosbecet, Gadolinium, Dimeglumine Gadopentetate, Gadoterate Meglumine, Gadoversetamide, Disodium Gadoxetate, Galactooligosaccharides, Galactooligosaccharides, Galantamine, Galantamine CR, Galeterone, Gallium Maltolate, Gallium Nitrate, Gallopamil, Gambo Acid, Gamma-Linolenic Acid, Gammatocopherol, Ganaxolone, Ganciclovir, Ganciclovir Phosphate, Ganetespib, Ganirelix Acetate, GanstigmineGarenoxacin, garlic, gatifloxacin, GCS-100, G-CSF, gedatricib, gefitinib, gelatin, gerofcin, gelpart, gemcaben, gemcitabine, gemcitabine elaidate, gemcitabine prodrug, gemfibrozil, gemifloxacin, gemigliptin, gemigliptin tartaric acid, gemzar, Genera, genistein, genistein + decitabine, gentamicin, gentamicin sulfate, gentamicin, gepiron, gepotidacin, gestodone, gestodone, Gestrinone, gilteritinib, gimatecan, gimeracil, ginkgo biloba, ginkgolide meglumine, ginsenoside Rg3, ginsenoside Rd, ginkobilova, ginkgodisadenafil, gibinostat, GKT-831, glatiramer acetate, glecaprevir, gresatinib glycolate, glibenclamide, gliclazide, glimepiride, glinide, glipizide, glitazone, GLP-1 analog, glucagon-like peptide-1, glucocorticoid, glucocorticoid, glucosamine, glucosamine Hydrochloride, glucose, gluphosphamide, glutamic acid, glutamine, glutathione, glycerin, glycerol, glycerol phenyl butyrate, glycoprotein IIb / IIIa inhibitor, glycopyrrolate, glycopyrrolate, glycopyrronium bromide, glycopyrronium tosylate, glycopyrronium bromide, glycyrlysine, gliminox, GM1, GM-CT-01, GnRH antagonist, sodium gold thiosulfate, golotimod, golbatinib tartrate, gonadotopine, gonadoto Ropin, Gonadotropin, Goodberry Probiotics, Goserelin Acetate, Goshazingan, Gosogliptin, gp100, GPO-VirZ30, Granisetron, Granotapide, Grazoprevir, Glepafloxacin, Griseofulvin, GR-MD-02, GSK-2269557, GSK-2330672, GSK-2339345, Guaifenesin, Guanethidine, Guanafacin, Gydiflusing, Gusperimus Trihydrochloride, GWP42004, Dicerol, Gynostemma pentapyllum, H2 Blocker, Haldol, Halofdinon, Haloperidol, Haloperidol Decanoate, Halothan, Hangeshacinto, HBW, HE-3286, Helon, Helico DR, Heliox,Heme alginate, hemin, hemoximer, heparin, sodium heparin, Her-2 / neu, heroin, hexachlorophene, hexaminolevulinic acid hydrochloride, Hextend, HF-0220, hibiscus subdalipid, hydrotarsid, himantan, histamine dihydrochloride, homatropin, honey, hpFSH, HPP-404, HQK-1004, huachansu, Huai Qi Huang, human chorionic gonadotrophin, huperzine A, Hyabest J, hyaluronic acid, sodium hyaluronate, hyaluronic acid, hyaluronic acid hydrogel, hydralazine, hydrochloric acid, hydrochlorothiazide, hydrochlorothiazide tablets, hydrochlorthiazide, hydrocodone, hydrocodone Vitarate, hydrocodone / acetaminophen, hydrocortisone, hydrocortisone sodium succinate, hydrocodone-17-butyrate, hydrocorton, hydrogel, hydrogen peroxide, hydromorphine, hydromorphone, hydromorphone hydrochloride, hydroquinidine, hydroquinone, hydroxocobalamin, hydroxycarbamide, hydroxychloroquine, hydroxydaunorubicin, hydroxyethyl starch, hydroxyethyl starch solution, hydroxyethylpropyl methylcellulose powder, hydroxymethyl butyrate, hydroxynortriptyline, hydroxyprogesterone caproate, hydroxypropylcellulose, hydroxytryptophan, hydroxyurea, hydroxyzine, helastan, hyalenic acid combination Substitute, hyoscin butyl bromide, hyoscin hydrobromide, hyoscin N-butyl bromide, hyoscyamine sulfate, high-pressure bupivacaine, hypericin, St. John's wort, hyperosmolar glucose, hypertonic saline, hypromellose, ibandronate, ibandronic acid, iverogasto, iverogasto N, IBH-B, ibipinabant, ivodutant, ivopamine, ibruci Ibdilast, ibuprofen, ibutamoren mesylate, ibuterol, ibutylide, icaritine, icodextrin, icofungipen, icosubtate, icosapent, icosapent ethyl, icosapent ethyl ester, icotinib hydrochloride, idalopyridine, idarubicin, idazoxane, IdB-1016, idebenone, idelalisib, idoxuridine,Hydrabiotaparinax sodium, hydranoxyl, iferanserin, ifetroban, ifetroban sodium, IFN-α2b, ifosfamide, iguratimod, IHBG-10, IL-2, iraprazole, ilepatril, iloperidone, iloprost, iloprost betadex clathrate, imagabalin, imatinib, ImCOOH, imeglimin, imexon, imidafenacin, imidap Lil, Imiglitazar, Imipenem, Imipramine, Imiquimod, Imisopasem manganese, IMO-2125, Impitapide, Incyclinide, Indacaterol, Indacaterol acetate, Indacaterol maleate, Indacaterol xinafoate, Indantadol, Indapamide, Indapamide SR, Indeglitazar, Indinavir, Indinolfort, Indipron, Indisetron, Indisram, Indisram, Indium In 111 anti-CD66 monoclonal antibody BW250 / 183, indocyanine green, indomethacin, indolamin, industrial nitric oxide, rice calcitol, INF-alpha, infigratinib, infliximab, ingavirin, ingenol mebutate, inhaled sodium nitrite, iniparib, injectable progestin, inosine, inosine pranobex, inositol, INS-1, insulin, insulin glargine, insulin NPH, intepirdin, interferon, interferon alpha, interferon alpha-2a, interferon alpha-2b, interferon beta-1a, interferon beta-1b, interleukin, interleukin-2, interferon-6, intertemumab, Intracel, intranasal ketamine, intravenous immunoglobulin, intravenous minocycline, iodine, iodine I-131, iodine 131 Ethiozide Oil, Iodine I 131 Monoclonal Antibody BC8, Iodine Coloring Agent, Iodixanol, Iohexol, Iopamidol, Iopromide, Ipragliflozin, Ipratropium, Ipratropium Bromide, IPX-159, IPX-231, Irbesartan, Irinotecan, Irinotecan Hydrochloride, Irinotecan Sulfonate, Ilofluben, Iron, Iron Folic Acid, Iron Hydroxide Polymaltose, Iron Oxides,Iron protein succinate, iron solution, iron sucrose, iron supplement, Ilostat, ylsogladine maleate, IRX-5183, ISA-51, isobuconazonium chloride / sulfate, Iscar, Iseganan, Isovid, Ocaloric Diet, isocarboxazide, isoflavone, isoflavone, isoflurane, isosorbide-S, isoniazid, isoniazid, isoprinosine, isopropyl alcohol, isopropyl unoprostone, isoproterenol, isoquercetin, isosorbide dinitrate, isosorbide mononitrate, isosorbide-5-mononnitrate, isosulfan blue, isotretinoin, isovaleramide, ispinecib, ispronicline, isradipine, islapafant, istaloxime, itacitinib, itasetron, itopride hydrochloride, itraconazole, itriglumide, Ivabradine, Ivabradine hydrochloride, Ibaftol, Ivermectin, Ixabepyrone, Isazomib citrate, Jinfukan, JNJ-56914845, Joverin, Josamycin, Jugransregia extract, Juvidex, Kamikihito, Kanamycin, Kaba, KD-018, Kefurin, Kenalog, Kenalog-10, Ketamine, Ketamine hydrochloride, Ketanserin, Ketoconazole, Ketoprofen, Keto Lorac, ketrolactromethamine, ketotifen, KLH, Crestin, KRX-0402, KT6-971, KW-2450, KW-2478, KWA-0711, KX2-391, L9NC, labetalol, labradimil, lasidipine, rasidofilstrong, lacosamide, lactated Ringer's solution, lactic acid, lactin-V, lactitol, lactic acid bacteria, lactic acid bacteria, lactic acid bacteria KS400, Lactobacillus Lactobacillus casei rhamnosus, Lactobacillus delbrueckii, Lactobacillus paracasei F19, Lactobacillus paracasei LP-33, Lactobacillus lactarose, Lactobacillus plantarum 299v, Lactobacillus reuteri, Lactobacillus lafutidine, L-alanosine, lamivudine, lamotridine, landiolol, lanisemin, laninamivir octanoate, lanikidal, lanoconazole,Lanopeptene, Lamperizone, Lansoprazole, Lanthanum carbonate, Lapaquistat, Lapatinib, Lakinimod, L-arginine, Laromustine, Laropiprant, Larotaxel, L-ascorbic acid, Rasmiditane, Lasofoxifen, L-asparaginase, Latanoprost, Latanoprost, Latanoprostembunod, Latrepyridine, Lauri, L-carnitine, lazabemid, LC-150444, L-carnitine, L-citrulline, LCL-161, Lcr-35, L-dopa, Recoboa, Recozotan SR, Redelforin, Ledipasvir, Lefamurin, Leflunomide, Lemborexant, Remteporfin, Lenalidomide, Lenograstim, Lentinan, Lentinan virus, Vatinib lenmesylate, LEO-80122, L-epinephrine, Lelcanidipine, Relsivirin, Recinurad, Lesogavelan, Restaurtinib, Retaxaban, Leteprinim, Letermovir, Letrozole, Leucine Leucaverin, leucovorin, leucovorin calcium, leucafferesis, leukotriene B4 (LTB4), leproleride, leproleline acetate, levalbuterol, levalbuterol hydrochloride, levamfetamine, levamisol, levamlodipine, levamlodipine besylate, levetiracetam, levobupivacaine, levosalbutamol hydrochloride, levocarnitine, levocetirizine, levocetirizine dihydrochloride, levodopa, levofloxacin, levofolinate, levogestrel, levocetoconazole, levoleucovorin, levomequitazine, lev Milnacipran, levonorgestrel, levofencinone hydrochloride, levofanol, levosalbutamol, levocimendan, levosulpiride, levothyroxine, levothyroxine sodium, levotophysopam, lexibrin, lexipaphant, L-folic acid, L-glutamine, LH-RH agonist, riafensin, riaozol, ribifem, ricarbazepine, licochalcone A, lycopheron, lidocaine, lidocaine chlorhydrate, lidocaine-prilocaine, rifibrol, refitegrass, lignocaine, LIK-066, li Maprost, Limtop, Linagliptin, Linaprazan, Lincomycin, Linezolid, Linifanib, Linoleic acid, Linopristin, Lincitinib, Liothyronine, Liothyronine sodium, Lipido, Lipiodol, Lipiodol-ethanol mixture, Liposin, LipoCol, Lipoic acid, Lipopolysaccharide, Liposomal amphotericin B, Liposomal cisplatin, Liposomal doxorubicin, Liposomal paclitaxel, Liposomal prostaglandin E-1, Liposomal vincristine, Reproca Depot, Lisabambrin hydrochloride, Lisdexamfetamine,Lisinopril, lysophilin, lyslide, lithium, lithium carbonate, citrate, lithium salt, litronsib, lixivaptan, L-leucovorin, L-leucyl-L-leucine methyl ester, L-NMMA, lobaplatin, robeglitazone, loberine, loberine sulfate, lobucavir, rodenafil carbonate, rodenosine, lofepramine, lofexidine, romibvir, lomustine, lomustine (CCNU), ronafarnib, ronaprisan, long-chain fatty acids, long-chain polyunsaturated fatty acids, ronidamin, loperamide, loperamide hydrotyrolide, loperamide Oxide, lopinavir, lopinavir / ritonavir, loratadine, lorazepam, locaserin, loredipron, lormetazepam, L-ornithine L-aspartate, loronoxicam, losartan, losartan potassium, rosigamon, rosmapimod, loteprednol etabonate, lovastatin, roxapine, loxoprofen, lubiprostone, lucanton, lucitanib hydrochloride, luliconazole, lumacafthol, lumateperone toluenesulfonate, lumefantrine, lumicitabine, luminespib, lumiracoxib, lunacalcipol, lurasidone, ruby Nectin, Lulotin, Lulutotecan, Luseogliflozin hydrate, Rustrombopag, Lutein, LY-2090314, LY-2623091, Livelid, Lycopene, Rimescycline, Linestrenol, Lysomutil, Makimorelin, Macitentan, Macrolid, Mafenide, Maphosphamide, MAGE-A1, MAG-EPA oil, Magnesium, Magnesium aluminum hydroxide, Magnesium carbonate, Magnesium chloride, Magnesium chloride hexahydrate, Magnesium citrate, Magnesium gluconate, Magnesium hydroxide Um, magnesium isoglycyrrhizinate, magnesium oxide, magnesium sulfate, magnesium valproate, malathion, managlinatodiaranetyl, mangafodipyr, manganese, manidipine, manidipine dihydrochloride, manitim, mannitol, mapracolate, maprotiline, malalixibat chloride, maraviroxine, maribavir, marijuana, marimastat, marberon, macilkast, macitinib, masopropyl, mabogulant, maxacalcitol, mazindol, MCC-135, MDR1, MDT-10013, mebendazole,Mebeberine, Mebeberine hydrochloride, Mecamillamine, Mechloretamine, Meclinate, Meclizine, Mecobalamin, Mecobalamin monohydrate, Medium-chain fatty acids, Medroxyprogesterone, Medroxyprogesterone acetate, Mefenamic acid, Mefloquine, Megestrol, Megestrol acetate, Meglumine antimonate, Melagatran, Melan A, Melalsoprole, Melatonin, Meldonium, Melphalan, Meloxicam, Merperone, Melphalan, Melphalan hydrochloride, Memantine hydrochloride, Menaquinone, Menaquinone-7, Menatetrenone, Me Nias, Menotropin, Menotropin, Menstrogol, Mepaclin, Meperidine, Mephalan, Mepivacaine, Mepivacaine Chlorhydrate, Mepivacaine Hydrochloride, Meprizine, MER-104, Melbaron, Mercaptamine, Mercaptamine Tartrate, Mercaptopuria, Merititabine, Melimepodiv, Meropenem, Mesalamine, Mesalazine, Mezna, Metadoxin, Metaforin, Metagrip, Metamisole, Metamisole Sodium, Metalaminol, Meteospasmil, Metformin, Metformin Lysinate, Metformin HCl, Metfo Lumin hydrochloride, metformin SR, methacholine, methadone, methadone hydrochloride, methamphetamine, metazolamide, methimazole, methionine, methocarbamol, methohexatal, methotrexate, methotrimeprazine, metoxsalen, methoxyflurane, methoxypsoralen, methylaminolevulinic acid hydrochloride, methylprednisolone, methylprednisolone acetate, methylcobalamin, methyldibromoglutaronitrile, methyldopa, methylene blue, methylnaltrexone bromide, methylphenidate, methylphenidate hydrochloride, methyl Rednisolone, methylprednisolone acetate, methylprednisolone acetate, sodium methylprednisolone succinate, methylprednisone, methylsamidolphan, methylsulfonylmethane, methyltetrahydrofolate, methylthioninium chloride, methylrosine, Metoves compounds, metocloperamide, metoclopramide, metrazone, metoprolol, metoprolol succinate, metoprolol tartrate, metoprolol XL, metronidazole, MetroGel, metronidazole, metronidazole cyclophosphamide, metyrapone,Mexiletine, Mexiletine Hydrochloride, Mexolyl SX, Mexolyl XL + Titanium Dioxide, Mianserin, Mibanpeter, MIBG, Miconazole, Miconazole Nitrate, Microalgae Oil, Microparticle Progesterone, Micronutrient Mixture, Midazolam, Midazolam Hydrochloride, Medium-Chain and Polyunsaturated Fatty Acids, Midodrin, Midostaurin, Mifepristone, Miflonide, Miglastat, Miglitol, Miglustat, Mirataxel, Milnacipran, Milrinone, Miltefosine, Milbeterol, Mimopezil, Minocycline, Minocycline Hydrochloride, Minodron Acids, minoxidil, mirabegron, miriplatin hydrate, mirodenafil, mirtazapine, misoprostol, mitemcinal, mitiglinide, mitogluzone, mitolactol, mitomycin, mitomycin C, mitokinone / mitokinol mixed redox mixture, mitotane, mitoxantrone, mibacrylium, mibacrylium chloride, mybobrin, mixed salt amphetamine, mizolastine, mizoribine, MK-0782, MK-0893, MK-2206, MK-7622, ​​MK-8457, MMF, mosetinostat hydrobromide, moclobemide, modafinil Moexipril, molidustat, morindon, morcidomin, molybdenum, momerotinib, mometasone, mometasone furoate, monolaurin, sodium glutamate, montanid ISA-51, montelukast, montelukast sodium, molacidine, morphine, morphine chloride, morphine glucuronide, morphine hydrochloride, morphine sulfate, mosapride, motesanib diphosphate, motexafingadolinium, motexafinlutetium, motlimod, moxidectin, moxifloxacin, moxifloxacin hydrochloride, moxonidine, MP-435, MSC-apceth-111, MT-102, mucaine, mucopolysaccharide, mucuna pruriens, multivitamin, muparfostat sodium, mupirosin, mulagritazal, mustine, Mycobacterium w, mycophenolic acid, mycophenolate mofetil, mycophenolate sodium, mycophenolic acid, mycostatin, mydriasert, myphenax, martus communis L, mitomycin, N2O-O2, nabilon, nabiximol, nabumethane, N-acetylcysteine, N-acetylcysteine, N-acetylcysteine,N-acetyl-L-cysteine, N-acetyl-p-aminophenol, NaCl, nadifloxacin, nadolol, nadropalin calcium, nafamostat, nafamostat mesylate, nafarelin, NaFeEDTA, naphthifine, naphthifine hydrochloride, naftopidil, nalbuffine, nalbuffine sebacate, naldemedine, nalfurafine, nalmefene, naloxegol, naloxone, naltrexone, naltrexone hydrochloride, naltrexone-poly(DL-lactide), nalzotan, namodenon, nandrolone, naosintong, napabucasin, naphthoquin, naproxinoid, naproxen, naproxen etemesyl, naproxen sodium, naratriptan, nalonapride, narrowband UVB, nasapake, nastrazepide calcium Natamycin, nateglinide, nabamepent, nabarixin, nabeglitazar, nabitoclax, N-chlorotaurine, nebentan, nebicapone, nebibolol, nebulizedmikacin, nebulized budesonide, nebusar, nedaplatin, nefazodone, nepadutant, nepafenac, nepicastat, neramexan, neratinib, neridronic acid, nerispiridine, netaludil, netylmycin, netivudine, netupitant, Neu-120, neutropin, nevirapine, niacin, niacinamide, Niacor, NIC5-15, nicardipine, nicergoline, nicolosamide, niconinamide, nicorandil, nicotinamide, nicotin, nicotinic acid, nifedipine, nifurtimo X, Nigella Sativa, Nicomycin Z, Nilotinib, Niltamide, Nilvadipine, Nimeslid, Nimodipine, Nimorazole, Nimotuzumab, Nimustine Hydrochloride, Nintedanib, Nipradilol, Niraparib, Nirogasestat, Nitazoxanide, Niticinone, Nitrazepam, Nitrendipine, Nitric Oxide, Nitrofluviprofen, Nitrofurantoin, Nitrogen Gas, Nitroglycerin, Nitroglycerin, Nitroprusside, Nitrous Oxide, Nitroxide, Nibokasan, Nizatidine, N-Lite, N-Monomethyl-L-Arginine, Novactin, Norethisterone, Norpitantium Acetate, Norethisterone Acetate Channel Blocker, Nonoxinol-9, Non-Preserved Latanoprost, Norelgestromine, Norepinephrine,Norhandrolone, norethindrone, norethindrone acetate, norethisterone, norethisterone acetate, norethisterone enanthate, norethlindone acetate, norfloxacin, norgestimate, nortriptylatin, nortriptyline, norlusodeoxycholic acid, noscapine, Nova, Bupivacaine, Novasoi, Novotax, NPC-18, NRTI, NS-2, NS-8, NTC-510A, NucleomaxX, Nucleoside e antiretroviral drug, NVB, Nistatin, Nistatin LF, Nikol, O2, Obatoclax, Obeticholic acid, Obicetrapib, Obinepitide, OBT, Ocaperidone, Osinapron, Octinoxate, Octocrylene, Octocrylene + Tinosolve, Octreotide acetate, Odalus, Ofloxacin, Ogremilast, Oglufanide disodium, Olanzapine, Olanzapine / Fluoxetine, Olaparib, Orsegepant, Oleic acid, Olepra, Oresoxime, Oryceridine, OligoG CF-5 / 20, olmesartan, olmesartan cilexetil, olmesartan hydrochloride, olprinone, olsalazine, oltipraz, omasetaxin mepesuccinate, omadacycline, omapatrillat, omamaligliptin, omaberoxolone, onbitasvir, omblabrin, omegatib mecavir, omega, omega-3, omega-3 carboxylic acid, omega-3 fatty acid, omega-3 polyun, omega-3 acid ethyl ester, omega-3 carboxylic acid, omega-6, omegaMAX, omegaben, omegabazole, omeprazole and bicarbonate, omeprazole sodium, o-methylphenidate, omidenepag propyl, omigapil, omaya reservoir, omri-hep-B, onarespib, oncovin, ondansetron, ondelopran, opicapon, opipra Mole, Opium, Fumagiline, Olanchinib, Orbofivan, Org-9426, Orilotimod, Oritabancin, Oristat, Ornithine Phenyl Acetate, Orteronel, Ortocaine, Orthosilicate, Orthostat, Orthostat-L, Orthobisc, Olbepitant, Oseltamivir, Osemozotan, OSI-632, Osilodrostat, Ospemifene, Osteodex, Osteonil, Otenabant, Oteracil Potassium, Oteseconazole, Othyronium Pamoate Oxyantel, Oxazepam, Oxycarbazepine, Sodium Glutathione Oxymax, Oxytriptan, Oxytropium Bromide, OX-NLA, Oxybuprocaine, Oxybutynin, Oxybutynin Chloride, Oxybutynin Hydrochloride, Oxycodone,Oxycodone CR, oxycodone oxysite, oxygen, oxymetazoline, oxymetazoline hydrochloride, oxymetholone, oxymorphone ER, oxymorphone IR, oxyprinol, oxytocin, ozagrel, ozagrel hydrochloride, ozanimod, onsenoxacin, P-276-00, P-53, PAC-14028, paclitaxel, paclitaxel polygrumex, paclitaxel-PM, pacritinib, pactimibe, paflamidine, pagoclone, paranocetron hydrochloride, palbociclib, paliphosphamide, paliperidone, paliperidone Don ER, paliperidone palmitate, pariroden, palivizumab, palonosetron, parovarotene, pamapimod, disodium pamidronate, PAN-90806, Panavir I, panobinostat, pantoprazole, pantothenic acid, pantovigor, papaverine, paquinimod, paracetamol, paldoprunix, parecoxib, paricalcitol, paritaprevir, pernaparin sodium, parogrelyl, paromomycin, paroxetine, paroxetine hydrochloride hemihydrate, paroxetine mesylate, parthenolide, Pasiflorine carnata, Paci Degib, pachylomer calcium, patpirone, pazopanib, pazufloxacin, pazufloxacin mesylate, PCI-24781, PCI-27483, PD-110843, PD-115934, pectin, pegamotecan, pegcantratinib, pegylated liposomal doxorubicin, PEITC, peritinib, peritrexol, perbiprofen, pemafibrate, pemetrexate, pemetrexetine, pemetrexetine disodium, pemirolast, pemirolast sodium, pemoline, penciclovir, penclomedine, penebiclidine hydrochloride Salt, penicillamine, penicillin, penicillin G, penicillin V, pentaerythritol tetranitrate, pentalite, pentamidine, pentamidine isethionate, pentazocine, pentobarbital, pentosan polysulfate sodium, pentostatin, pentotar, pentoxifylline, pentoxifylline, peramivir, perclozone, peretinoin, perflubron emulsion, perflutren lipid microsphere, pergolide, perhexylline, perifosin, periryl alcohol, perindopril, perindopril arginine,Permethrin, Perospirone, Perphenazine, Perdiinfotel, Pethidine, Pethidine Hydrochloride, PF-04447943, PF-05089771, PF-05175157, PF-3654746, PF-3654764, PF-4191834, PF-4531083, PF-4691502, PF-489791, PF-610355, PG-2, PGL-2001, PGP / BCRP Inhibitor, PH-797804, Phenelzine, Phenindione, Pheniramine Maleate, Phenobarbital, Phenobarbital Sodium, Pheno Xybenzamine, fenprocumone, fencerine, phentermine, phentolamine mesylate, phenylbutyrate, phenylephrine, phenylephrine hydrochloride, phenytoin, phloroglucinol, PHN-031, PHN-033, phosphate, phosphatidylserine, phosphatidylcholine, phosphatidylcholine-related naproxine, phosphatidylcholine-encapsulated ibuprofen, phospholax, phosphorus, photopheresis, physostigmine, phytate, phytonadione, phytosterols, pivocerod, pibrentasvir, piciva Nil, picridenoson, piclozotan, picoplatin, picotamide, piclorif, piclopodophyllin, picloriza, pictilisib, piraralisib, pilocarpine, pilocarpine hydrochloride, pilsicainide, PIM-447, pimacertive hydrochloride, pimavanserin, pimecrolimus, pimoziv, pimozide, pindolol, pinosembrin, pioglitazone, pioglitazone hydrochloride, pipemperone, piperacillin, piperacillin sodium, piperacillin-tazobactam, piperacin, piperacin phosphate, piperine, piracetam, pyrariatin Pirarubicin, pirenzepine, pirfenidone, pyribezil, pyridoxine, pyritramide, pyritrekim, pyromelatin, piroxicam, pitavastatin, pitavastatin calcium, pitrizant, pivamecillinum, pibmecillinum, pixantrone, PL-3994, plant sterols, platinum, prazomycin, preconalil, prelixafor, previtrexed, prinabulin, PLX-8394, PM-00104 / 50, PMI-001, PMK-N02RS1, pokapavir, polaprezinc, policosanol, polidocanol,Polyfeprosan 20 containing carmustine, polmacoxib, polyethylene glycol, polyethylene glycolated IL-2, polyethylene glycol citrate-simethicone, high molecular weight nanoparticle docetaxel, polymyxin B sulfate, polyphenone E, polysaccharide-K, polysorbate 80, polysporine, polytetrafluoroethylene, pomagleta domethionyl, pomalidomide, ponatinib, ponesimod, polactant alfa, porfimer sodium, porphyromycin, posaconazole, posizolide, posterizan acuto, potassium, potassium canrenoate, potassium chloride, potassium iodide, potassium nitrate, potassium perchlorate, povidone, povidone iodine, pozaniclin, poziotinib, PPA Lux 680, PPA-904, PPD-10558, PPI, PR-104, Prasinostat, Pradefovir, Prazigastat, Pralatrexate, Pralidoxime, Prarunacasan, Pramiconazole, Pramipexole, Pranlukast, Pranlukast hydrate, Plasteron, Prasugrel, Pravastatin, Praziquantel, Prazosin, Prednicarboate, Prednisolone, Prednisolone acetate, Prednisolone phosphate, Prednisolone sodium metazoate, Prednisolone sodium succinate, Prednisone, Pregabalin, Pregnenolone, Preradinant, Premarin, Prempro, Prenatal vitamin, Presatovir, Pretomanid, Prilocaine, Primaquine, Prinavelel, Prinomast, Priteri Viru, probenecid, probiotics, probucol, procainamide, procaine, procaine hydrochloride, procarbazine, procarbazine hydrochloride, procaterol, procaterol hydrochloride, prochlorperazine, procysteine, prodocosapentaenoic acid, proeicosapentaenoic acid, progesterone, progestin, progestogen, progestogen dienogest, proglumide, proguanil, proguanil hydrochloride, proralix, promethazine, prometra, promethorium, promisan, propacaine hydrochloride, propaketamol, propafenone, propafenone-SR, propranolol, proparacaine, propionyl-L-carnitine, propiverine, propranolol hydrochloride, propranolol, propranolol repro,Propranolol-Repro, Propranolol, Propranolol Hydrochloride, Propranolol LA, Propranolol XL, Propylthiouracil, Propifenazone, Prosolva, Prostaglandin, Prostaglandin-E2, Prostin, Proteros, Protriptyline, Provide Ectra, Proximetacaine, Proximetacaine Hydrochloride, PRS-211375, Purcalopride, Prulifloxacin, Pro Lurisol, Pulvanserin, PRX-3140, PRX-8066, PSD-508, Pseudoephedrine, Pseudoephedrine hydrochloride, Pseudoephedrine mannose-sensitive hemagglutinin (PA-MSHA), PSI-5004, PSI-938, Psilium powder, PTH, p-toluenesulfonamide, puerarin, puerarin sodium phosphate, pumocetrag, PVAC, PVP-ILH liposome, PX-12, PYN- 17. Pyrantel-oxantel, pyrazinamide, pyridostigmine bromide, pyridoxal, pyridoxamine dihydrochloride, pyridoxine, pyridoxine hydrochloride, pyrimethamine, pyrimethamine / sulfadoxine, pyrimethamine / sulfadoxine, pyronaridine, Q-301, QAV-680, kimbudan, quidton gluo, quercetin, ketiapine, ketiapine fumarate hydrochloride, ketiapine IR, quinacrine, quinacrine hydrochloride Golid, quinapril hydrochloride, quinifamide, quinidine, quinine, quinolone, quinupristin, xinostat, quizartinib dihydrochloride, R-112, labacfosazine, rabeprazole, rabeximod, ravesertib, racecadotril, lacloprid, radafaxine, radalbuvir, ladequinil, radezolid, rad-108, radioactive iodine, radioactive iodine, radiolabeled iodobenzamide, radiprodil, radium Ra 223 Dichloride, Radotinib, Lagaglitazar, Ralfinamide, Raloxifene, Raltegravir, Laritrexed, Ramatroban, Ramelteon, Ramipril, Ramosetron, Ranibizumab, Ranimustine, Lanirestat, Ranitidine, Lanolazine, Rapamycin, Rasagiline, Lasegulant, Rabidasvir Hydrochloride, Labconazole, Laxatolidine, Razpenem, RBP-8000, RBx-10017609, RDEA-806, RDP-58, Rebamipide, Rebaudioside A, Levimast, Leboxetine,Refametinib, modified diclofenac, modified mebendazole, rigadenoson, regorafenib, reglelol, lerebactam, lerenoprid, relugolix, remifentanil, remimazolam, remimazolam tosylate, remogliflozin etabonate, REN-1654, Lenzapride, repaglinide, Reparixin, Repinotan, Repajuremic acid, Repaondansetron, Resatorvid, Reserpine, Resiniferatoxin, Resminostat, Resoferon, Respifor, Respimat, Restilane, Restilane SubQ, Resveratrol, Letagliptin, Letapamulin, Letaspimycin, Retigavin, Retinoic acid, Retinoid, Retinol, Retosiban, Retrovir, Rebamilast, Lebefenacin, Lebexepride, Leviparin sodium, Lezatomidine, RF-07026, rFSH, RG-4929, RG-723 4, RG-7795, RGH-478, RGH-507, Rhenium-188-HEDP, Rhinox, RhuDex, Ribavirin, Ribociclib, Riboflavin, Ridaforolimus, Ridinirazole, Rifabutin, Rifaradil, Rifampicin, Rifampin, Rifamycin, Rifapentin, Rifaximin, Rigosertib sodium, Ricunsit, Lilapradib, Rilmenidine, Rilpivirine hydrochloride, Riluzole, Limakarib, Rimejepant, Rimexolone, Limonabant, Ringer's acetate, Ringer's lactate solution, Riociguat, Rifa hydrochloride Radiryl hydrate, risedronate sodium, risperidone, ritanserin, ritobegron ethyl hydrochloride, ritodrine, ritonavir, rituximab, rivaroxaban, rivastigmine, rivenprost, ribipancel sodium, riboglitazone, rizatriptan, RLP-068, RNF43, RNS-60, RO-4929097, RO-5036505, lovarzotan, rociletinib, rocuronium, rocuronium bromide, lofecoxib, loferon-A, loflumilast, lorapitant, loripram, lorofilin, lonacarelet, loni Cyclib, lonopterin, ropinirole, ropinirole hydrochloride, ropivacaine, lokinimex, rose bengal sodium, rosiglitazone, rosiglitazone maleate, rosiglitazone XR, rosiptol acetate, rostafloxin, rostaporfin, rosuvastatin, rosuvastatin calcium, rotigotine, lovatirelin, roxadustat, roxithromycin, RP-323, RPh-201, RPL-554, RPM-02 / 08, RQ-00000004, R-salbutamol sulfate, rubitecan, ruboxystaurin, rucaparib,Lucaparib camsilate, lucaparib phosphate, rufinamide, rupatadine, ruxolitinib, S-ketamine, S(+)-ketamine, S-1, S-111, S-38093, S-707106, SAB-378, sabarbicin, saccharomyces brardii, sacubitril, S-adenosylmethionine, SAF-312, saffron, safinamide, safotivant, sagopyrone, salbutamol, salbutamol HFA, salbutamol sulfate salicylic acid, salicylic acid + benzoic acid, salidroside, physiological saline, salilacib, salmon calcitonin, monkey Salate, salubrin, samarium (153Sm) lexidonam, samatasvir, samidolfan, S-amlodipine centitulate, sapacitabine, sapanicertib, sapitinib, sapropterin, sapropterin dihydrochloride, saquinavir, SAR-110894, salakatinib, salecycline, saredutant, salglamostim, salizotan hydrochloride, saroglitazal, sarpogrelate hydrochloride, satavaptan, satraplatin, saxagliptin, SB-773812, SBP-002, SC-49483, SCH-002063, SCH-497 079, SCH-900776, Cysandrus phenantella extract, scopolamine, SCY-078, SDX-101, secnidazole, cedritia rosmarin, segesterone acetate, ceradelper L-lysine dihydrate, selegiline, selegiline hydrochloride, selenium, selenium sulfide, selenomethionine, cerepressin, celetracetam, selexipag, sericlib, selinexor, sericistat, serodenosone, ceroncertib, cerozoc, selumetinib, cerlanpanel, semagacestat, semapimod, semacsanib, sembragiline, senicapok , senna, sensorkaine, Centra PM, theocalcitol, separatentronium bromide, cepetaprost, cepranolon, septin, SER-150-DN, selenorepens, cergliflozin etabonate, cerulopitant, serotonin reuptake inhibitor, serotonin / norepinephrine reuptake inhibitor, cerutaconazole, certindol, sertraline, S-ethylisothiourea diethyl phosphate, cetyluton, setipipant, S-etodrug, cetrobuvir, sevelamer carbonate, sevelamer hydrochloride, sevoflurane, cevofuran,Sebuparin sodium, SG-2000, Shanfan Uji decoction, SHP-465, Sibrafiban, Sibutramine, Sildenafil, Sildenafil citrate, Silexan, Silibin, Silibinin dihydrogencosinate, Silymarin, Silodosin, Silver nanoparticles, Silver nitrate, Silver sulfadiazine, Silibin, Silymarin, Simenepagisopropyl, Simeprevir, Simethicone, Simethicone, Symvastatin, Simbalo, Siponimod, Sirolimus, Sitafloxacin, Sitagliptin, Sitaquin, Sitaxentan, Sitosterol, Sibe Restat, Cibichen, S-Ketamine, SKP-1052, SK-PC-B70M, SLx-4090, SMANCS, S-Methionyl-L-Citrulline, Sumiragenin, SMP-028, SN-38, SNX-5422, Soderglytazar, Sodium, Sodium 4-Phenylate, Sodium Ascorbate, Sodium Benzoate, Sodium Bicarbonate, Sodium Butyrate, Sodium Carboxymethylcellulose, Sodium Chloride, Sodium Cromoglycate, Sodium Citrate, Sodium Dichloroacetate, Sodium Ferric Gluconate Complex Sodium fluoride, sodium folate, sodium fusidate, sodium vitreous luteinate, sodium hydroxide, sodium hypochlorite, sodium ketolac, sodium lactate, sodium nitrate, sodium nitrite, sodium nitrite, sodium oxybate, sodium phenylacetate, sodium phenylbutyrate, sodium phenylbutyrate, sodium sodium butyrate, sodium phosphate, sodium picosulfate hydrate, sodium polystyrene sulfonate, sodium prasterone sulfate, sodium pyruvate, sodium stibogluconate, sodium sulfide, tetra Sodium decyl sulfate, sodium thiopental, sodium thiosulfate, sodium valproate, sodium fluoride, sofinicline, sofosbuvir, sofosbuvir + daclatasvir, sofpyronium bromide, sorabegron, sorcitinib, sordesam, solifenacin, sorisromycin, soluble ferric pyrophosphate citrate, solvazinc, somatostatin, sonedenoson, sonidegib, sonolisib, sorafenib, soraprazan, sorbitol, sorivudine, sotagliflozin, sotalol, sotrastaurin, sobapreva, soy isoflavones,Sparcetan, spbrutinib, spiramycin, spironolactone, sPLA2 inhibitor, SQ-109, squalamine, SR-T100, SSS, ST-101, STA-4783, standard bicarbonate, standard pulmonary peritoneum, tin compounds, stansoporfin, statins, stabudin, stearidonic acid, S-tenatoprazole, sterofandin, steroids, steroids, stiripentol, stopine, Streptococcus faecium, Streptococcus thermophilus, streptomycin, streptozosin, strontium chloride Sr 89, Strontium malonate, Strontium ranerate, Strontium-89, STW-5, STX-107, SU, SU-101, SU-14813, Succimer, Succinic acid, Succinylcholine, Sucralphat, Sucralose, Sucropheric oxyhydroxylate, Sufenta, Sufentanil, Sufentanil citrate, Sufentanil, Sugammadex, Sulbactam, Sulfadoxin sodium, Sulf Adoxin, sulfadoxine, sulfadoxine + pyrimethamine, sulfarene-pyrimethamine, sulfamethoxazole, sulfasalazine, sulfate solution, sulfonylurea, sulfonylurea, sulfonylurea, sulforapan, sulindac, suldoxide, sulopenim, sulopenim etozadroquil, sulfacetoamide sodium, sulpadoxin, sulfacetamide sodium, sulfadoxine, methox Sazole, sulfonylurea, sulpiride, sultamicillin, smanilol, sumatriptan, sumatriptan succinate, SUN-0597, SUN-1334H, sunitinib, suplatast silate, suramin, suramin sodium, surinabant, stezolid, suzamethonium, SYI-2074, symbiotic, synbiotics, SYNSORB-Pk, synthetic hypericin, T / S, T-122 5, T-2000, T3, tacalcitol, tasejinarin, tacrine, tacrolimus, tacrolium hezal, tadalafil, tadequinyl alfa, tafamidis, tafenoquin, tafluprost, tafoquiparin sodium, TAK-715, TAK-783, talabostat, taladegib, tarampanel, talaporfin, taralazole, tarazoparib, talc, TALL-104, taeumapimod, tarnetant,Talsacridine fumarate, taltirelin, tamibarotene, tamoxifen, tamsulosin, tamsulosin hydrochloride, tandospirone, tandutinib, tanespimycin, tanomast, tandicertib, tapentadol, taprenepag, taradial, tarafenacin, taranaban, tarenfluvir, taribavirin hydrochloride, talikidal, tarloxtinib bromide, tacidotin HCl, tasimelton, tasisla Taspoglutido, Tascinimod, Taurolidine, Tauroursodeoxycholic acid, Tababolol, Tabirmid, Taxol, Taxus, Tazarotene, Tazobactam, TC-2403, TC-3, TCM-606F, Tebipenim pivoxil, Tecadenoson, Tecalcet, Tecaphalin, Tecastemisole, Technetium bicitate, Technetium ethalphoratide, Technetium Tc 99m Chilmanocept, Technetium Tc 99m trophorastat, technetium-99, tecobilimat, tedathioxetine, tedisamil, tedizolide phosphate, tegapher, tegaserod, tegralinad chloride, tegobuvir, teicoplanin, telaprevir, telapristone acetate, teratinib, terbivudine, terkagepant, telithromycin, telmisartan, telotoritol etiplate, temazepam, temocapril, tenmosilin, tenmoporfin, temozolomide, temsirolimus, tenapanol, teneligliptin, teniposide, tenofovir, tenofovir alafenamide, tenofovir alafenamide fumarate, tenofovir disoproxil fumarate, tenofovir exalidex, tenofobi Lu / emtricitabine, tenoxicam, teprenone, telameprocol, terazosin, terbinafine, terbinafine acetate gel, terbinafine hydrochloride, tervogrel, terbutaline, terbutaline sulfate, terconazole, terguride, teriparatide, terlipressin acetate, tertroban, tesaglitazal, tesetaxel, tesemiliphen, tesofensin, tetagen, testosterone, cypionate testosterone, testosterone enanthate, testosterone undecanoate, tetanus toxoid, tetomirast, tetrabenazine, tetracaine, tetracaine hydrochloride, tetracycline, tetracycline HCl, tetrahydrobiopterin,Tetrahydrocannabinol, tetrathiomolybdate, tetrodin, tezacaftol, tezacitabine, tezanpanel, tezosentan, TG-100-115, TG-100801, thalidomide, THC, theophylamine, theophylline, theophylline SR, ceracrumin, thiamine, thioamine hydrochloride, thiazide, thiazide, Diuretics, thiazolidinedionic acid, thioguanine, thiocorticoside, thioctic acid, thioguanine, thiopental, thiopental sodium, thiopental, thioridazine, thiotepa, thiothixen, THR-0921, THR-4109, thrombin, thrombin microcapsules, thymokutonan, thymoglobulin, thyroxine, thiagabin, thianaptine, tiapride, tiborone, ticaglecol, ticlopidine, tideglucib, tigecycline, tirapertin, tyralginine acetate, tildronate disodium, timapiprant, ticodal, tinodonic acid, timolol, timolol, timolol maleate, tinidazole, tinzaparin sodium, thiopronin, thioto Ropium, tiotropium bromide, tipiracil hydrochloride, tipranavir, tirapazamine, tiracemtib, tililazad, tirofiban, tirofiban hydrochloride, titanium dioxide, TIVA, tivantinib, tivozanib, tizanidine, TMC-114 / RTV, TMC-310911, TMC-647055, TNF-alpha, TNP-470, tobramycin, tocladesine, tocofersolan, tocopherol, tocopherol, tocopheryl phosphate mixture (TPM), tofacitinib, tofimilast, tofogliflozin, tolcapone, tolvamer, tolnaphthat, tolperisone, tolterodine, tolterodine tartrate, tolvaptan, TOMM34, tonaversat, tonarin SG1000T FFA, Tonapophylline, Tonjinluo, Topiramate, Topiroxostat, Topotelan, Topotelan hydrochloride, Trazemide Torcetrapib, Trefolan, Toremifene, Tosedostat, Tosufloxacin, Tozadenant, Tozacertib, Trabectedin, Trabectome, Travodenoson, Tradipitant, Tramadol, Tramadol hydrochloride, Tramazoline, Trametinib, Tramiprosate, Trandolapril, Tranexamic acid, Tranilast, Transcrocetinate sodium, Tranylcypromine, Trastuzumab, Travoprost, Trazoprodil, Trazadone, Trazodone hydrochloride, TRC-101, Trehalose, Trelagliptin succinate, Trelanserin, Treosulfan, Treprostinil, Treprostinyl diolamine, Tretinoin, Triamcinolone, Triamcinolone acetonide,Triamcinolone hexacetonide, Triapine, Triatec HCT, Triaz, Triazabilin, Triazolam, Tribenzimidine, Trichlormethiazide, Trichlorothiazide, Trisirivine, Tridolgosyl, Trientine, Trientine hydrochloride, Triphalotene, Trifluoperazine, Trifluridine, Triflusal, Triheptanoin, Trihexyphenidyl, Triiodothyronine, Trilostane, Trimebutine, Trimebutine trimethobenzuamide, Trimethoprim, Trimethop Limu-sulfa, Trimethrexate, Trimipramine, Trimipramine maleate, Trinitrate, Triomune, Triplixum, Tripotasium disitrate bismutate, Triptorelin Willfordi, Triptorelin acetate, Triptorelin pamoate, Trisodium citrate dihydrate, Trivatropisetron, Trospium chloride, Troxacitabine, Troxipide, Tornature, Triptorelin-EDTA, TS-022, TTK, TTP-05 4, TTP-399, TTP-435, TTP-889, TTX-9401, Tsucaresol, Tusidinostat, Tulobuterol, TV-46763, TV-5010, TY-51924, Tirenol, TZD, TZP-102, Ubidecarenone, Ubiquinol, Hubrogepant, UCA-001, UCA-002, UCN-01, Udenafil, UFT, UFT / LV, UFUR, UISH-001, UK-390957, UK-432097, UK-447841, Urimorelin, Urinasta Chin, Ulipristal, Urobetasol, Urodesine, Umeclidinium, Umeclidinium Bromide, Umirolimus, Unfractionated Heparin, Uniphylline, Upadacitinib, Upamostat, Uprifosbuvir, Uprosertib, UR-906, Uracil, Urapidil, Urea, Uridine, Uridine Triacetate, URLC10, Ursodeoxycholic Acid, Ursolic Acid, Ulticadiosea, Bocistapis, UTD-1, Utrogestan, V-116517, V-158866, V-404, VA -111913, babicaserin, babolbactam, vadadastat, badimethan, badiprost, VAK-694, valacyclovir, valacyclovir, valbenazine, valdecoxib, valerian extract, valganciclovir, barium,Valnivudine hydrochloride, Valnoctamide stereoisomer, Valnoctamide stereoisomer, Baloma, Valrubicin, Valsartan, Valsartan trisodium heptaminehydrate, Valspodar, Baltrucitabine, Vancomycin, Vancomycin hydrochloride, Vandetanib, Vaniprevir, Vanoxerine, Bapendavir, Bapitadine, Vardenafil hydrochloride, Varenicline, Valesprazib, Valesprazib methyl, Baritinib, Vasopressin, Batalanib, Batiquinone, VDC-2008, Vecuronium, Vecuronium bromide, Vedroprevir, VEGFR1, VEGFR1 peptide, VEGFR1-A02- 770, VEGFR2 peptide, VEGFR2-derived HLA-A0201, Beliflapon, Beliparib, Verneperit, Belpatasvir, Bercetrag, Vemurafenib, Venetoclax, Venlafaxine, Venlafaxine hydrochloride, Venlafaxine XR, Bepoloxamer, Verapamil, Verapamil hydrochloride, Versilnon, Verdiperstat, Verisigwat, Verinurad, Vernacarrant, Vernacarrant hydrochloride, Verteporfin, Belbrin, Versaphon, Vertex, Besatrimod, Besnarinone, Bestipitant, Vibegron, Bicriviroc, Vidoca, Vidofludimus, Vidupiplant, Bigabatrin, Vitamin D, Vilanterol, Vilanterol triphenate, Viraprisan, Virazodone, Vildagliptin, Vinblastine, Vinblastine sulfate, Vincristine Vincristine, vincristine sulfate, vindesine, vinflunin, vinorelbine, vinorelbine dipropyl, vinpocetine, vintafolide, vipadenant, bisodesib, vitusertive, vitamin B12, Vitamac, vitamin A, vitamin B, vitamin B1, vitamin B12, vitamin B-12, vitamin B2, vitamin B6, vitamin B-6, vitamin C, vitamin D, vitamin D2, vitamin D3, vitamin E, vitamin E succinate derivative, vitamin K, vitamin K1, vitamin B1, vitamin B12, vitamin B2, vitamin B6, vitreosolve, biucid, bizomitin, vohopitant, voglibose, borasertib, borinanserin, vonoprazan fumarate, borapaxer, voriconazole, vorinostat, voltioxetine hydrobromide, vosaroxine, boxylaprevir, vocalisib,VP-101, VRx-3996, VSL-3, VVZ-149, VX-105, VX-135, VX-702, VX-710, Warfarin, Warfarin, Warfarin sodium, Water, WF-10, WH-1, Wobendym, WX-554, Zariproden, Xemirofiban, Xenon, Xiang-sha-liu-jun decoction, xiaoqinglong, Xibrom, Xilei-San, Ximelagatran, Xi'anping, XL-139, XP-21279, Xylitol, Xylometazoline, Y-39983, Yaraferon, Yonkenafil, Yttrium cribatuzumab tetraxetan, Yttrium Y90 anti-CD66 monoclonal antibody BW Examples of small molecule drugs include, but are not limited to, 250 / 183, zavofloxacin, zafirlukast, zalcitabine, zaleplon, zaltoprofen, zanamivir, ZD-6126, zeaxanthin, dibotentan, diclonapine, zidovudine, dileton, zinc, zinc acetate, zinc oxide, zinc picolinate, zinc sulfate, zinc supplements. Other examples include, but are not limited to, small molecule drugs such as Zinkasforte, dinthionine, ziprasidone, zofenopril, zoledronic acid, zoliflodacin, zolmitriptan, zolpidem, zolpidem tartrate, zometa, zonanpanel, zonisamide, zonisamide SR, zopicron, zoskaider, zotarolimus, zotepine, zucapsaicin, zuclopentisol, and zuretinol acetate.

[0223] In some embodiments, the manipulated platelet cargo described herein is a small molecule, such as one from the CeMM Library of Unique Drugs (CLOUD), as shown in Licciardello et al., Nat Chem Biol; Vol. 13, pages 781-780 (2017), which is incorporated herein by reference in its entirety. For example, small molecules include pinacidyl, altretamine, pipobromane, uracil mustard, trioxalene, plicamycin, ambenonium, edrophonium, hexafluorenium, oxytriphylline, albutamine, guanabenz, mephentermine, methoxamine, phenylpropanolamine, protochlor, tetratrazoline, trazoline, betanidine, ergoloid, oxyprenolol, penbutrol, phentolamine, propiomazine, thiethylperazine, fomepizol, triamterene, stanozolol, dromostanolone, ethylestrenol, fluoxymesterone, methyltestosterone, deserupidine, quinapril, rescinnamine, spirapril, testolactone, ethionamide, sulfamethazole, sulfacitin, sulfamerazine, sulfamethazine, sulfamethizol, sulfafenazole, sulfapyridine, sulfathiazole, sulfi Squizazole, sulfoxone, cefmenoxide, amdinocillin, azurocillin, bacampicillin, carbenicillin, cephalothin, cefditoren, cefonisid, cefolanide, cefotiam, cefpyramide, cefradin, ceftizoxime, cephaloglysin, cefapillin, cyclacillin, hetacillin, loracalbef, methicillin, meslocillin, moxalactam, nafcillin, ticalcillin, capromycin, demeclocycline, jiris Romycin, Metacycline, Oxytetracycline, Spectinomycin, Troleandmycin, Biomycin, Enoxacin, Novobiocin, Alatrofloxacin, Synoxacin, Lomefloxacin, Enalidixic acid, Sparfloxacin, Trovafloxacin, Acetohydroxamic acid, Marinol, Ethoxzolamide, Acetohexamide, Fenoprofen, Oxyfenbutazone, Carprofen, Oxaprozin, Phenylbutazone,Tolmetine, Meclophenamic Acid, Methylergonovine, Acetofenadine, Carphenazine, Chlorprothixen, Mesolidazine, Triflupromazine, Promazine, Benzfetamine, Phentrazine, Chlorotrianicene, Estrone, Mestranol, Polyestradiol, Kinestrol, Cortisone, Fluprednisolone, Meprednisone, Paramethasone, Oxamnicine, Azatadine, Bromodifenhydramine, Buclidine, Carbinoxamine, Clofedianol, Dexbrompheniramine, Diphenylpyraline, Mepyramine, Mesdiradin, Trimeprazine Tripelennamine, Triprolidine, Romidepsin, Pramidone, Butabarbital, Chlormezanone, Plurazepam, Butabarbital, Chlormezanone, Flurazepam Quazepam, Secobarbital, Tarbutal, Thiamylal, Gamma-hydroxybutyrate, Memantine, Triclofos, Piperazine Hexahydrate, Deoxycorticosterone Pivalate, Pargiline, Carbachol, Oxyphenonium Bromide, Anisotropin, Cridinium, Cyclimin, Dicyclomine, Difemanil, Etopropazine, Fesoterodine, Hexosylium Isopropamide, mepenzolate, methantheline, methylscopolamine, methixene, orphenadrine, oxyphencycline, procyclidine, propantheline, tridihexetyl, throspium, decametonium, pentolinium tartrate, cisatoricrium besylate, succinylcholine chloride, doxacrium, galamine, methocrine, pancuronium, pipecuronium, rapacuronium, tubocurarine, guanadrel, fendimethrazine, anilelysine, diphenoxine, diphenoxylate, levometasil, oxymorph Propoxyfen, levallorphan, methylnaltrexone, (+ / -)-sulfinpyrazone, pamidronic acid, risedronate, tildronate, clofibrate, diphylline, inamlinetin, vardenafil, etinodiol, hydroxyprogesterone, norethinodrel, ulipristal acetate, carboprost, etretinate, methisergidone, chlorphentermine, acetyldigitoxin, deslanoside, chlorpropamide, trazamide, tolbutamide, chlorothiazide, cyclothiazide, hydroflumethiazide,This may include, but is not limited to, polythiazide, quinetadine, 5-fluorouracil, dextrothyroxine, methirosine, rimantadine, adefovir, anisindione, dicumarol, nisoldipine, trimetadine, bepridil, paramethadine, bretylium tosylate, mephenytoin, benzonate, etotoin, indecinide, mollicidine, phenasemide, tokainide, pyrvinium chloride dihydrate, halophanthrine, metaxalon, diphenidol, mebutamate, chlorphenesin, fenximide, thiabendazole, benzquinamide, piperacetadine, eschlorbinol, and etinamate.

[0224] IV. Production In some embodiments, the manipulated platelets described herein can be produced using the technique described in Ito et al. (Cell, 174(3):636-648.e18, 2018), which is incorporated herein in whole by reference. Ito provides a method for producing platelets from iPSC precursors on a clinical scale. Turbulence has been observed to activate platelet biosynthesis for clinical-scale ex vivo production of platelets from human induced pluripotent stem cells (iPSCs) (ibid.). iPSCs derived from the immortalized megakaryocyte precursor cell line (imMKCL) were combined with the soluble factors insulin-like growth factor-binding protein 2 (IGFBP2), macrophage migration inhibitory factor (MIF), and nardilysin combatase (NRDC) in a bioreactor where the physical parameters of turbulent energy and shear stress are controlled (ibid.).10 11 Platelet production exceeding [a certain value] was observed (ibid.). The platelets were observed to function similarly to those derived from the donor (ibid.).

[0225] In certain embodiments of the present invention as described herein, imMKCL can be established by introducing the cancer-derived MYC(c-MYC) / Polycomb ring finger oncoproto-gene (BMI-1) and BCL2 1-like 1 (BCL-XL) genes into iPSCs using a lentivirus. Additional genes may be introduced or deleted to produce edited megakaryocytes, in fact, even platelet-specific promoters have been previously characterized. These genes provide inducible gene expression in the presence of drugs such as doxorubicin (DOX). imMKCL can be cryopreserved until culture is desired. Megakaryocyte proliferation is stimulated by contacting the cell line with the drug, resulting in the expression of the inserted genes. The drug is removed to stop gene expression and allow platelet production.

[0226] Current FDA-approved regulations for the storage of platelets for transfusion require storage at 22°C and use within 6 days. By reference, the entire report, Slichter et al., “Treatment of Bleeding in Severely Thrombocytopenic Patients with Transfusion of Dimethyl Sulfoxide (DMSO) Cryopreserved Platelets (CPP) Is Safe—Report of a Phase 1 Dose Escalation Safety Trial,” Blood, 2016, is incorporated here, assuming that platelets can be cryopreserved for 2 years when frozen with DMSO. Following a positive Phase 1 trial, Phase 2 and Phase 3 trials are ongoing. Infusion of up to 3 units of cryopreserved platelets (CPP) into patients with severe thrombocytopenia and active bleeding appeared to be “safe and without evidence of thrombotic complications, despite CPP having a procoagulative phenotype due to the cryopreservation process.” Therefore, cryopreserved platelets may have the effect of stabilizing, reducing, or stopping bleeding in patients with thrombocytopenia, as measured using the World Health Organization (WHO) bleeding grade. No evidence was found to undermine the hypothesis that cryopreserved platelets used for non-coagulation purposes are as effective as platelets stored according to current FDA regulations.

[0227] V. Pharmaceutical Compositions This instruction further includes a pharmaceutical composition comprising one or more of the manipulated platelets of the present invention and, optionally, at least one pharmaceutically acceptable excipient or inactive component. Furthermore, the pharmaceutical may include the therapeutic delivery system described herein.

[0228] As used herein, the term “pharmaceutically acceptable composition” means one or more of the engineered platelets described herein, or preparations of their pharmaceutically acceptable salts, with the optional use of other chemical components such as physiologically appropriate carriers and excipients.

[0229] The term "excipient" or "inactive ingredient" refers to an inert or inactive substance added to a pharmaceutical composition to facilitate the administration of a compound. Non-limiting examples of such inactive ingredients are disclosed below in the formulations of this specification.

[0230] In some embodiments, the composition is administered to a human, a human patient or subject. For the purposes of the present disclosure, the phrase "active ingredient" generally refers to any one or more of the engineered platelets delivered as described herein.

[0231] The description of the pharmaceutical compositions provided herein is primarily directed to pharmaceutical compositions suitable for administration to humans, but those skilled in the art will understand that such compositions are generally suitable for administration to other animals, such as non-human animals, such as non-human mammals. Subjects to which administration of the pharmaceutical composition is contemplated include non-human mammals, including but not limited to agricultural animals such as cows, horses, chickens and pigs, domestic animals such as cats and dogs, or research animals such as mice, rats, rabbits, dogs and non-human primates.

[0232] The pharmaceutical compositions according to the invention can be prepared, packaged, and / or sold in bulk as a single unit dose and / or as multiple single unit doses. As used herein, a "unit dose" is an individual quantity of a pharmaceutical composition containing a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient administered to the subject and / or a convenient fraction of such a dose, such as half or one third of such a dose.

[0233] The relative amounts of the active ingredient, pharmaceutically acceptable excipients or inactive ingredients, and / or any additional ingredients in the pharmaceutical compositions according to the invention vary according to the identity, size, and / or condition of the subject being treated, and further according to the route by which the composition is administered. By way of example, the composition may contain from 0.1% to 100%, such as from 0.5 to 50%, from 1 to 30%, from 5 to 80%, at least 80% (w / w) of the active ingredient.

[0234] The effectiveness of treatment or improvement of a disease can be evaluated, for example, by measuring disease progression, disease remission, symptom severity, pain reduction, quality of life, the dosage of drugs required to maintain the treatment effect, the level of disease markers, or other measurable parameters suitable for a given disease being treated or prevented. Monitoring the effectiveness of treatment or prevention by measuring any one of such parameters, or any combination of parameters, is within the ability of one of ordinary skill in the art. In connection with administration of the compositions of the present invention, for example, being "effective" against cancer means that administration in a clinically appropriate manner results in improvement of symptoms, cure, reduction of disease burden, reduction of tumor mass or cell number, extension of lifespan, improvement of quality of life, or other effects generally recognized as positive by physicians skilled in the treatment of a particular type of cancer, etc., and results in a beneficial effect in at least a statistically significant proportion of patients.

[0235] A treatment or preventive effect is demonstrated by a statistically significant improvement in one or more parameters of the disease state, or by the absence of deterioration or onset of symptoms otherwise expected. As an example, a preferred change of at least 10%, preferably at least 20%, 30%, 40%, 50% or more in a measurable parameter of the disease may indicate an effective treatment. The effectiveness of a given composition or formulation of the present invention can also be determined using an experimental animal model of a given disease known in the art. When using an experimental animal model, if a statistically significant change is observed, the effectiveness of the treatment is demonstrated.

[0236] A. Therapeutic Delivery System Various embodiments of the invention herein provide a non-thrombogenic therapeutic delivery system comprising at least one engineered platelet, also referred to as a SYNLET™ therapeutic delivery system.

[0237] The SYNLET® therapeutic delivery system can be produced using the megakaryocyte / platelet production strategies described herein. Precursors can be sequentially edited without affecting safety, creating the possibility of designing a fully synthesized network. Immunogenic cargo can be encapsulated within platelets and subsequently released at the target site. Antigen sensitivity is controlled via a loading difference between CPR-ITAM and the inhibitor CPR-ITIM.

[0238] 1. Cancer treatment In some embodiments, the engineered platelets described herein may be used to treat solid tumors. In the field of immuno-oncology, engineered platelets may be modified to be activated by antigen-specific T cells, potentially upregulating their function to eliminate tumors expressing defined novel antigens. Conversely, antigen-specific T cells mediating autoimmune diseases may be targeted for destruction with defined antigens known in various common diseases such as Hashimoto's thyroiditis, type 1 diabetes, and multiple sclerosis.

[0239] In one embodiment, the targeting portion of CPR can recognize CD19 to deliver chemotherapeutic agents locally. CD19 is a well-known B cell surface molecule that, upon activation of the B cell receptor, promotes B cell antigen receptor-induced signaling and expansion of the B cell population. CD19 is widely expressed in both normal and neoplastic B cells. B cell-derived malignancies, such as chronic lymphocytic leukemia, acute lymphoblastic leukemia, and many non-Hodgkin lymphomas, frequently retain CD19 expression. This nearly universal expression and specificity to single cell lineages makes CD19 an attractive target for immunotherapy.

[0240] In some embodiments, the targeting portion of the CPR may recognize tumor-specific antigens (TSAs), such as neoplastic antigens expressed only by tumor cells due to gene mutations or transcriptional changes that alter the protein-coding sequence, and thus may create novel, exogenous antigens. Genetic changes result from gene substitutions, insertions, deletions, or other genetic alterations of native congener proteins (i.e., molecules expressed in normal cells).

[0241] For example, a tumor may be starved by platelets, leading to clot formation and a cessation of blood supply. Alternatively, CPR can target tumors by expressing cargo or toxins directed to alpha granules that mediate the local delivery of therapeutic agents. In additional embodiments, CPR may target tumors by expressing antibodies directed to alpha granules, such as checkpoint inhibitors, to increase anti-tumor immunity.

[0242] The engineered platelets described herein can be engineered to kill cancer cells. For example, CD19 has been targeted to TRAIL-expressing platelets for the treatment of cancerous B-cell leukemia (BCL). CD19-targeted CAR-T cells show great clinical promise compared to BCL. TNF superfamily members (TRAIL) and Fas ligand (FASL) have been shown to induce BCL death via apoptosis upon CD40 stimulation (see Dicker et al. “Fas-ligand (CD178) and TRAIL synergistically induce apoptosis of CD40-activated chronic lymphocytic leukemia B cells”. Blood, 2005, which is incorporated herein by reference in its entirety). CD40L is spontaneously exposed to activated platelets (see Henn et al. "CD40 ligand on activated platelets triggers an inflammatory reaction of endothelial cells". Nature, 1998, which is incorporated herein by reference in its entirety), and therefore may activate the FASL / TRAIL-dependent cell death pathway when bound to BCL, and FASL is spontaneously exposed to activated platelets (see Schleicher et al. "Platelets induce apoptosis via membrane-bound FasL". Blood, 2015, which is incorporated herein by reference in its entirety). Platelets expressing TRAIL have been used to reduce prostate cancer metastasis in mice (see Li et al. "Genetic engineering of platelets to neutralize circulating tumor cells". Journal of Controlled Release, 2016, which is incorporated herein by reference in its entirety).In one embodiment, resting platelets presenting CD19-single-chain variable fragment (scFv)-ITAM and containing TRAIL, CD40L, and FASL ligands are activated by the binding of CD19 and CD19-scFv-ITAM on B cells. Upon activation, TRAIL, CD40L, and FASL are presented on the platelet surface. Platelet-induced death of leukemia cells is mediated by CD40L binding to the CD40 receptor on B cells, activating the FASL / TRAIL-dependent cell death pathway.

[0243] In certain embodiments, platelets can be engineered to direct the proliferation of novel antigen-specific T cells in vivo. Novel antigens are present in many human tumors and can be identified computer-aidedly. Ex vivo proliferation and reinjection of T cells results in the death of the targeted tumor. Immune checkpoint inhibition allows T cells to kill tumors expressing the novel antigen (however, non-specificity can lead to serious side effects). Megakaryocytes can be loaded with MHC class 1 molecules containing exogenous peptides, which can then be transferred to platelets. Novel antigens may be expressed in megakaryocytes, and the MHC class 1-ITAM fusion protein can stimulate checkpoint inhibitors. This enables in vivo proliferation of novel antigen-specific T cells. For example, platelets can be engineered to express MHC1-nascent antigen-ITAM. Both engineered platelets and T cells are activated by the interaction of MHC1-nascent antigen-ITAM with the novel antigen-specific T cell receptor (TCR). Upon activation, cytotoxic T lymphocyte-associated protein 4 (CTLA4) and programmed cell death 1 (PD-1) are presented on the platelet surface, interacting with CTLA4 inhibitors (CTLA4i) and PD-1 inhibitors (PD-1i) on T cells, respectively. Maximum T cell activation and proliferation are achieved by blocking checkpoints.

[0244] 2. Treatment of autoimmune diseases In some embodiments, the manipulated platelets described herein may be used to treat autoimmune conditions. At least 81 autoimmune diseases have been identified in humans, of which 45 are associated with autoantigens, and 36 autoantigens are tissue-specific (see Hayter, et al., Autoimmunity Reviews 11 (2012) 754–765, which is incorporated herein by reference in its entirety). Autoimmune diseases were defined as disorders in which 1) a specific adaptive immune response is directed toward an affected organ or tissue, 2) autoreactive T cells and / or autoantibodies are present in the affected organ or tissue, 3) autoreactive T cells and / or autoantibodies can transmit the disease to a healthy individual or animal, 4) immunity to autoantigens induces the disease in animal models, and 5) elimination or suppression of the autoimmune response prevents disease progression and even improves clinical symptoms. An additional criterion considered for the definition was that antibody binding interferes with the function of the autoantigen (ibid.). Self-tolerance checkpoints have also been identified at each stage of lymphocyte development and activation (ibid.).

[0245] In some embodiments, the manipulated platelets described herein may be used to suppress autoantigen-specific T cells to treat autoimmune diseases. In some embodiments, the CPR in the manipulated platelets may include regions specific to tissues associated with autoantigens. For example, the tissues are selected from the group consisting of adipose tissue, adrenal glands, ascites, bladder, blood, bone, bone marrow, brain, cervix, connective tissue, ear, embryonic tissue, esophagus, eye, heart, intestine, kidney, larynx, liver, lung, lymph, lymph nodes, mammary glands, mouth, muscle, nerve, ovary, pancreas, parathyroid gland, pharynx, pituitary gland, placenta, prostate, salivary gland, skin, stomach, testes, thymus, thyroid gland, tonsil, trachea, umbilical cord, uterus, blood vessels, and spleen.

[0246] Table 12 shows the molecular and / or tissue targets of a non-exclusive list of neurological autoimmune disorders by Hayter, et al. [Table 12]

[0247] Table 13 shows the molecular targets and / or tissue targets of a non-exhaustive list of endocrine autoimmune disorders by Hayter, et al. [Table 13]

[0248] Table 14 shows the molecular targets and / or tissue targets of a non-exhaustive list of gastrointestinal autoimmune disorders by Hayter, et al. [Table 14]

[0249] Table 15 shows the molecular targets and / or tissue targets of a non-exhaustive list of hematopoietic autoimmune disorders by Hayter, et al. [Table 15]

[0250] Table 16 shows the molecular targets and / or tissue targets of a non-exhaustive list of musculoskeletal autoimmune disorders by Hayter, et al. [Table 16]

[0251] Table 17 shows the molecular targets and / or tissue targets of a non-exhaustive list of skin and mucosal autoimmune disorders by Hayter, et al. [Table 17]

[0252] Table 18 shows the molecular targets and / or tissue targets of a non-exhaustive list of skin autoimmune disorders by Hayter, et al. [Table 18]

[0253] Table 19 shows the molecular and / or tissue targets of a non-exclusive list of cardiovascular autoimmune disorders by Hayter, et al. [Table 19]

[0254] Table 20 shows the molecular and / or tissue targets of a non-exclusive list of other autoimmune disorders by Hayter, et al. [Table 20]

[0255] In some embodiments, the engineered platelets include a CPR having a region recognized by disease-mediating autoreactive T cells. For example, the CPR includes an epitope from molecular targets listed in Tables 12-20 loaded onto an MHC-ITAM fusion to directly target autoreactive T cells. The engineered platelets can be loaded with cytotoxic or immunosuppressive proteins or antibodies that are released upon platelet activation.

[0256] For example, some cases of type 1 diabetes (T1DM) are characterized by T cells specific to certain insulin peptides. Therefore, using an MHC1-ITAM receptor fusion protein together with an autoimmune-driven peptide in platelets designed to release immunosuppressive factors results in T cell-specific immunosuppression. Many other potential options regarding exposure to IL-2 receptors (IL-2R) that compete with IL-2, release of TGF-β1 or IL-10, and MHC1-ITAM activation are regulatory T(T) reg It mediates immunosuppression, similar to other cells.

[0257] In some embodiments, engineered platelets containing CPRs having major histocompatibility complex (MHC) class I or II are used to treat autoimmune diseases. T cells expressing chimeric antigen receptors (CARs) containing the MHC ligand of a pathogenic T cell receptor as the antigen-binding domain of the CAR have been previously shown to be effective in treating type 1 diabetes (T1D) (see Perez et al., Immunology, 143, 609-617, which is incorporated herein by reference in its entirety). In T1D, autoreactive CD8 and CD4 T cells selectively destroy insulin-producing B cells of the pancreas (ibid.). Some MHC-II restriction peptides recognized by autoreactive cells have been observed to originate from insulin / preproinsulin, islet-specific glucose-6-phosphatase catalytic subunit-related proteins, glutamate decarboxylases 65 and 67, heat shock proteins 60 and 70, insulinoma-related protein 2, zinc transporter ZnT8, islet amyloid polypeptide, chromogranin A, and other autoantigens (ibid.). Accordingly, in some embodiments, the engineered platelets described herein contain CPRs having ligands or fragments thereof that interact with autoreactive cells to destroy them.

[0258] Several autoimmune conditions (e.g., autoimmune thyroiditis, pemphigus vulgaris) are driven by antibody-dependent processes. CAR-T cells were previously created to target B cells that mediate autoimmunity (see Ellebrecht et al. Science 2016). The engineered platelets described herein, expressing the autoantigen-ITAM CPR, can kill B cells involved in the production of autoantibodies that drive autoimmunity. Activation of engineered platelets with B cell binding can enable the death of specific B cells. An "AND" gate can be engineered to allow only the activation of engineered platelets in response to autoantigen-B cell receptor (BCR) and CD19 binding, thereby increasing the specificity of delivery of cargo or toxins loaded onto the platelets.

[0259] 3. Treatment of infectious diseases Platelets are also involved in the clearance of bacterial infections and constitute a key component of the innate immune response; therefore, the SYNLET® therapeutic delivery system can be used to treat drug-resistant infections.

[0260] VI. Medication and Management The present invention provides a method comprising administering one or more compositions for immunotherapy to a subject in need thereof. These can be administered to the subject in any amount and via any route of administration that is effective in preventing or treating clinical conditions such as cancer, infections, and other immunodeficiency diseases.

[0261] The pharmaceutical compositions according to the present invention are typically formulated in dosage unit form for ease of administration and uniformity of dosage. However, it will be understood that the total daily dose of the compositions of the present invention may be determined by the attending physician within the bounds of sound medical judgment. The level of a particular therapeutic or prophylactic effective dose for a particular patient will depend on a variety of factors, including the disorder being treated and its severity, the activity of the particular compound used, the particular composition used, the patient's age, weight, general health status, sex and diet, the timing of administration of the particular compound used, the route of administration, previous or concurrent therapeutic interventions and excretion rates, the duration of treatment, any drugs used in combination with or concurrently with the particular compound used, and similar factors well known in the medical field.

[0262] The term “effective dose” refers to the amount of an active ingredient necessary to prevent or alleviate at least one sign or symptom of a particular disease and / or condition, and relates to a composition in sufficient quantity to provide the desired effect. Thus, the term “therapeutic effective dose” refers to the amount of an active ingredient or composition containing an active ingredient sufficient to promote a particular effect when administered to a typical subject. Effective doses also include amounts sufficient to prevent or delay the onset of disease symptoms, alter the course of disease symptoms (e.g., slow the progression of disease symptoms), or reverse disease symptoms. It is understood that, in any given case, a suitable “effective dose” can be determined by a person skilled in the art using routine experiments.

[0263] The pharmaceutical, diagnostic, or prophylactic compositions of the present invention can be administered to a subject using any amount and route of administration that is effective in preventing, treating, managing, or diagnosing a disease, disorder, and / or condition. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the disease, the specific composition, its mode of administration, its mode of activity, etc. The subject may be human, mammal, or animal. The compositions according to the present invention are typically formulated in unit dosage forms for ease of administration and uniformity of dosage. However, it will be understood that the total daily dose of the compositions of the present invention may be determined by the attending physician within the bounds of sound medical judgment. The level of a specific therapeutically effective dose, prophylactically effective dose, or appropriate diagnostic dose for a particular individual will depend on a variety of factors, including the disorder and its severity being treated, the activity of the specific payload used, the specific composition used, the patient's age, weight, general health, sex and diet, time of administration, and route of administration, duration of treatment, drugs used in combination with or concurrently with the active ingredient, and similar factors well known in the medical field.

[0264] In certain embodiments, the pharmaceutical composition according to the present invention may be administered at a dosage level sufficient to deliver approximately 0.01 mg / kg to approximately 100 mg / kg, approximately 0.01 mg / kg to approximately 0.05 mg / kg, approximately 0.05 mg / kg to approximately 0.5 mg / kg, approximately 0.01 mg / kg to approximately 50 mg / kg, approximately 0.1 mg / kg to approximately 40 mg / kg, approximately 0.5 mg / kg to approximately 30 mg / kg, approximately 0.01 mg / kg to approximately 10 mg / kg, approximately 0.1 mg / kg to approximately 10 mg / kg, or approximately 1 mg / kg to approximately 25 mg / kg of the target body weight per day, once or more, in order to obtain the desired therapeutic, diagnostic, or prophylactic effect.

[0265] The desired dose of the pharmaceutical composition described herein may be delivered once, three times daily, twice daily, once daily, every other day, every three days, every week, every two weeks, every three weeks, or every four weeks. In certain embodiments, the desired dose may be delivered using multiple doses (e.g., two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, or more doses). When multiple doses are used, divided dosing regimens as described herein may be used. As used herein, “divided dose” means dividing a “single unit dose” or total daily dose into two or more doses, e.g., two or more “single unit doses.” As used herein, “single unit dose” is the dose of any therapeutic agent administered in a single dose / at once / through a single route / at a single point of contact, i.e., in a single dosing event.

[0266] In some embodiments, depending on the properties of the artificial platelets, they can be introduced into a host organism, such as a mammal, by a wide variety of methods, including injection, transfusion, infusion, local injection, or transplantation. In some embodiments, the manipulated platelets of the present invention can be introduced into the site of a tumor. The number of manipulated platelets used will depend on many circumstances, the purpose of introduction, the lifespan of the manipulated platelets, the protocol used, e.g., the number of doses, the capacity of the manipulated platelets, multiplication, etc. The manipulated platelets may be contained in a physiologically acceptable culture medium.

[0267] In some embodiments, the manipulated platelets of the present invention may be administered in multiple doses to subjects with a disease or condition. The administration generally results in improvement of one or more symptoms of cancer or a clinical condition, and / or treats or prevents cancer or its clinical condition or symptoms.

[0268] The pharmaceutical composition comprising the manipulated platelets of the present invention can be administered via any route to achieve therapeutically effective results.

[0269] These routes include: intestinal (intestines), gastrointestinal, epidural (intradural), oral (through the mouth), percutaneous, intradural, intracerebral (in the cerebrum), intraventricular (intraventricular), subcutaneous (applied to the skin), intracutaneous (into the skin itself), subcutaneous (under the skin), nasal administration (through the nose), intravenous (intravenous), intravenous bolus, intravenous infusion, intraarterial (into the artery), intramuscular (into the muscle), intracranial (into the heart), intraosseous injection (into the bone marrow), intrathecal (into the spinal canal), intraperitoneal (injection or injection into the peritoneum), intranasal injection, intravitreous (through the eye), intravenous injection (into the lesion), intracavitary (into the base of the penis), vaginal administration, intrauterine, extraamniotic administration, and transcatheter Skin (diffusion from intact skin throughout the body), transmucosal (diffusion through mucous membranes), transvaginal, injectable (by snorting), sublingual, sublabial, enema, eye drops (superconjunctival), eye drops, auricle (inside or via the ear), cheek (towards the cheek), conjunctiva, skin, dental (into one or more teeth), electroosmosis, intracervical, intranasal sinuses, intratracheal, extracorporeal, hemodialysis, infiltration, interstitium, intraperitoneal, intraamniotic, intraarticular, intrabile duct, intrabronchial, intrasacral, intracartilage (intracartilage), caudal (caudal equine), cisterna magna (intracartilage macrocephalic cyst), intracornea (intracornea), intracornea, intracoronary artery (intracoronary artery), intracavitary (intracavitary space of the corpus cavernosum of the penis), intervertebral space Intraplate (intravertebral disc), intratubule (intraductal duct), intraduodenum (intraduodenum), intradura (intrathoracic or subdural), intraepidermis (up to the epidermis), intraesophagus (up to the esophagus), intrastoma (intrastomach), intragingiva (intragingival duct), intraileum (distal part of the small intestine), intralesional lesion (direct introduction or to localized lesion), intraluminal lumen (intratubular lumen), intralymph (intralymph), intramedullary (intramedullary cavity of bone), intrameninges (intrameninges), intramyocardium (intramyocardium), intraocular (intraocular), intraovary (intraovarian), intrapericardium (intrapericardium), intrapleura (intrapleura), intraprostate (intraprostate), lung (intralung or its bronchi), nasal cavity, testis (intratestis), intramedullary cavity (any level of the brain-spinal axis) Intravascular (intracerebrospinal fluid), intrathoracic (intrathoracic cavity), intratubular (intratubular cavity of organs), intratumoral (intratum), intratympanic (in the auricle), intravascular (intravascular or intravascular space), ventricle (intraventricular cavity), iontophoresis (by electric current causing ions of a soluble salt to move into body tissues), irrigation (to immerse or wash out open wounds or body cavities), larynx (directly into the larynx), nasogastric (from the nose to the stomach), occlusive dressing techniques (local administration followed by covering with a dressing that occludes the area), ophthalmology (into the external eye), oropharynx (directly into the mouth and larynx), parenteral, percutaneous, periarticular, peridural, perineurial,Examples of interventions include, but are not limited to, periodontal, rectal, respiratory (intravascular by oral or nasal inhalation for local or systemic effects), posterior orbital (behind the cerebral cortex or behind the eyeball), intramyocardium (in myocardial aging), soft tissue, subarachnoid, subconjunctival, submucosal, local, transplacental (through or across the placenta), transtracheal (through the tracheal wall), transtympanic (cross or through the tympanic cavity), ureter (to the ureter), urethra (to the urethra), vagina, caudal block, diagnostic, nerve block, biliary perfusion, cardiac perfusion, photopheresis, or spinal.

[0270] VII.Definition Throughout this specification, the characteristics or functions of the compositions of this disclosure are disclosed in groups or scopes. This disclosure is particularly intended to include all individual subcombinations of members of such groups and scopes. The following is a non-limiting list of term definitions.

[0271] As used herein, the term “antigen” is defined as a molecule that elicits an immune response when it is introduced into a target or produced by a target, such as a tumor antigen resulting from cancer development itself. This immune response may include antibody production, or activation of cells with specific immunological capabilities, such as cytotoxic T lymphocytes or T helper cells, or both.

[0272] As used herein, the terms “approximately” or “about” refer to a value similar to the reference value stated when applied to one or more values ​​of interest. In certain embodiments, unless otherwise specified or evident from the context, the terms “approximately” or “about” refer to a range of values ​​that are less than or equal to (greater than or less than) 25, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 in either direction of the reference value stated (except when such a number exceeds 100).

[0273] As used herein, the terms “related,” “conjugated,” “linked,” “attached,” and “tethered” mean, when used in relation to two or more parts, that the parts are physically related or linked to one or more additional parts, either directly or through one or more additional parts acting as linkers, to form a structure that is sufficiently stable so that the parts remain physically linked under the conditions in which the structure is used, for example, under physiological conditions. “Linked” does not necessarily have to be strictly via a direct covalent chemical bond. “Related” may also suggest that the ionic or hydrogen bond, or hybridization-based connectivity, is sufficiently stable so that the entities remain physically related.

[0274] As used herein, the term “cancer” refers to a broad group of diseases characterized by the uncontrolled growth of abnormal cells within the body. Unregulated cell division and growth lead to the formation of malignant tumors that invade adjacent tissues and eventually metastasize to distant parts of the body via the lymphatic system or bloodstream.

[0275] As used herein, the term “cytokine” refers to a family of small, soluble factors with pleiotropic expression capabilities produced by many cell types that can influence and regulate the function of the immune system.

[0276] As used herein, the term “delivery” means the act or method of delivering a compound, substance, entity, part, cargo, or payload. “Delivery agent” means any agent that at least partially facilitates the in vivo delivery of one or more substances (including, but not limited to, the compounds and / or compositions of the present invention) to cells, subjects, or other biological system cells.

[0277] When used herein, embodiments of the invention described herein are "operated" if they are designed to have features or properties, whether structural or chemical, that are derived from the starting point, wild type, or natural molecule.

[0278] As used herein, “expression” of a nucleic acid sequence means one or more of the following: (1) generation of an RNA template from a DNA sequence (e.g., by transcription), (2) processing of an RNA transcript (e.g., by splicing, editing, 5' cap formation, and / or 3' end processing), (3) translation of RNA into a polypeptide or protein, (4) folding of a polypeptide or protein, or (5) post-translational modification of a polypeptide or protein.

[0279] As used herein, “Preparation” includes at least the compounds and / or compositions of the present invention and a delivery agent.

[0280] As used herein, “fragment” refers to a part of a protein. For example, a protein fragment may include a polypeptide obtained by digesting a full-length protein. In some embodiments, a protein fragment contains at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 150, 200, or 250 or more amino acids. In some embodiments, an antibody fragment contains a part of an antibody.

[0281] As used herein, the term “immune cell” refers to any cell of the immune system derived from hematopoietic stem cells in the bone marrow that gives rise to two main lineages: myeloid progenitor cells (which give rise to myeloid cells such as monocytes, macrophages, dendritic cells, megakaryocytes, and granulocytes) and lymphoid progenitor cells (which give rise to lymphoid cells such as T cells, B cells, and natural killer (NK) cells). Exemplary immune system cells include CD4+ T cells, CD8+ T cells, CD4-CD8-double-negative T cells, Tγδ cells, Tαβ cells, regulatory T cells, natural killer cells, and dendritic cells. Macrophages and dendritic cells are sometimes referred to as “antigen-presenting cells” or “APCs,” in which major histocompatibility complex (MHC) receptors on the surface of APCs, which form complexes with peptides, interact with the TCR on the surface of T cells.

[0282] As used herein, the term “in vitro” refers to events occurring in an artificial environment, such as a test tube or reaction vessel, cell culture, or petri dish, rather than within a living organism (animal, plant, or microorganism).

[0283] As used herein, the term "in vivo" refers to an event occurring within a living organism (for example, an animal, plant, or microorganism, or their cells or tissues).

[0284] As used herein, “linker” or “targeting domain” refers to a portion of a chimeric platelet receptor that recognizes and binds to a desired antigen.

[0285] As used herein, “checkpoint factor” refers to any part or molecule whose function acts at a junction of a process. For example, a checkpoint protein, ligand, or receptor may function to halt or accelerate the cell cycle.

[0286] As used herein, the term “messenger RNA” (mRNA) refers to any polynucleotide that codes for a polypeptide of interest and can be translated to produce the encoded polypeptide of interest in vitro, in vivo, in situ, or ex vivo.

[0287] As used herein, the term “mutation” refers to a change and / or alteration. In some embodiments, a mutation may be a change and / or alteration of a protein (including peptides and polypeptides) and / or nucleic acid (including polynucleotides). In some embodiments, a mutation includes a change and / or alteration of a protein and / or nucleic acid sequence. Such a change and / or alteration may include the addition, substitution and / or deletion of one or more amino acids (in the case of proteins and / or peptides) and / or nucleotides (in the case of nucleic acids and / or polynucleotides, e.g., polynucleotides). In some embodiments in which a mutation includes the addition and / or substitution of amino acids and / or nucleotides, such addition and / or substitution may include one or more amino acid and / or nucleotide residues and may include modified amino acids and / or nucleotides. The resulting construct, molecule, or sequence of a mutation, change, or alteration may be referred to herein as a variant.

[0288] Where used herein, the term “novel antigen” refers to tumor antigens that are present in tumor cells but not in normal cells and do not induce deletion of their homologous antigen-specific T cells in the thymus (i.e., central tolerance). These neo-tumor antigens may provide “external” signals similar to those of pathogens to induce the effective immune response required for cancer immunotherapy. Novel antigens may be limited to specific tumors. Novel antigens are peptides / proteins with missense mutations (missense neo-antigens) or novel peptides with long, completely novel amino acid stretches from novel open reading frames (neoORFs). In some tumors, they can be generated in some tumors by out-of-frame insertions or deletions (due to defects in DNA mismatch repair causing microsatellite instability), gene fusions, read-through mutations of stop codons, or translation of improperly spliced ​​RNA (see Saeterdal et al., Proc Natl Acad Sci USA, 2001, 98:13255-13260, which is incorporated herein by reference in its entirety).

[0289] As used herein, the term “pharmaceutically acceptable excipient” refers to any component other than the activator (e.g., as described herein) that is present in a pharmaceutical composition and is substantially non-toxic and non-inflammatory in the subject. In some embodiments, a pharmaceutically acceptable excipient is a vehicle capable of suspending and / or dissolving the activator. Examples of excipients include antifouling agents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (pigments), emulsifiers, fillers (diluents), film-forming agents or coatings, flavors, fragrances, flow accelerators (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspenders or dispersants, sweeteners, and hydration water. Examples of excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, dibasic calcium phosphate, calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropylcellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0290] A pharmaceutically acceptable salt of a compound described herein is a form of the disclosed compound in which the acidic or base portion is in salt form (e.g., produced by reacting a free base group with a suitable organic acid). Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acidic salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. Typical acid addition salts include acetates, adipices, alginates, ascorbicates, aspartates, benzenesulfons, benzoates, bisulfates, borates, butyrates, camphorates, camphor sulfons, citrates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfons, fumarates, glucoheptons, glycerophosphates, hemisulfons, heptons, hexanoates, hydrobroms, hydrochlorides, hydroiodides, and 2-hydroxyethanesulfonesulfones. Examples include acid salts, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malons, methanesulfons, 2-naphthalenesulfons, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectins, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propions, stearates, succinates, sulfates, tartrates, thiocyanates, toluenesulfons, undecanoates, and valersates. Typical alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, as well as ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine, but are not limited to these; non-toxic ammonium, quaternary ammonium, and amine cations are also included. Examples of pharmaceutically acceptable salts include conventional non-toxic salts from non-toxic inorganic or organic acids. In some embodiments, pharmaceutically acceptable salts are prepared from a parent compound containing a basic or acidic moiety by conventional chemical methods.Generally, such salts can be prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of a suitable base or acid in water or an organic solvent, or in a mixture of the two (generally, a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile is preferred). A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, Pharmaceutical Salts: Properties, Selection, and Use, PHStahl and CGWermuth (eds.), Wiley-VCH, 2008, and Berge et al., Journal of Pharmaceutical Science, 66, 1-19 (1977), each of which is incorporated herein by reference in whole.

[0291] As used herein, the terms “subject” or “patient” refer to any organism to which the compositions according to the present invention may be administered, for example, for experimental, diagnostic, preventive, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, ra...

Claims

1. A modified megakaryocyte or its precursor that can reduce thrombus formation ability and / or produce platelets with reduced thrombus formation ability, at least GPIb / V / IX and GPVI (GP6), ITGA2B, CLEC2, Integrin s α ＩＩｂ β ３ , α ２ β １ , α ５ β １ and α ６ β １ One gene encoding a protein involved in the recognition of the primary stimulus for thrombus formation, selected from the group consisting of or from the group consisting of GPVI and ITGA2B, Par1, Par4, P2Y12, GPIb / V / IX, thromboxane receptor (TBXA2R), P2Y1, P2X1, and integrin α ＩＩｂ β ３ a protein involved in the recognition of secondary mediators of thrombosis, selected from the group consisting of, or selected from the group consisting of Par1, Par4, and P2Y12, and a gene encoding such a protein One gene encoding a protein involved in the release of secondary mediators of thrombus formation, selected from the group consisting of Cox1, HPS, and thromboxane A synthase (TBXAS1), or selected from the group consisting of Cox1 and HPS. Manipulated megakaryocytes or their precursors, including the destruction or deletion of a megakaryocyte.

2. The following genes: The manipulated megakaryocyte or precursor according to claim 1, wherein each of GPVI, ITGA2B, Par1, Par4, P2Y12, Cox1, and HPS is destroyed or deleted.

3. The platelets produced by the manipulated megakaryocytes or their precursors are a) Normally, it does not respond to endogenous stimuli that lead to blood clot formation, b) Not recruited by other activated platelets, and / or c) The manipulated megakaryocyte or precursor thereof according to claim 1 or 2, which, when activated, is unable to recruit and activate the patient's endogenous platelets.

4. The manipulated megakaryocyte or precursor according to any one of claims 1 to 3, wherein the disruption of the gene comprises selectively altering the expression of the gene with an RNA interference construct (RNAi), small interfering RNA (siRNA), microRNA (miRNA), or short hairpin RNA (shRNA).

5. The manipulated megakaryocyte or precursor according to any one of claims 1 to 4, wherein the disruption or deletion is a complete disruption or deletion of the entire gene.

6. The manipulated megakaryocyte or precursor according to any one of claims 1 to 5, wherein the destruction or deletion is a disruption of the function of the protein.

7. It is non-immunogenic and can be selected at will. a) The function of endogenous MHC class 1 is destroyed and / or b) The β2 microglobulin gene is disrupted. The manipulated megakaryocyte or its precursor according to any one of claims 1 to 6.

8. The megakaryocyte or its precursor contains a chimeric platelet receptor, and the chimeric platelet receptor is a) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, b) A heterologous targeting domain that recognizes and binds to a target, The manipulated megakaryocyte or precursor according to any one of claims 1 to 7, comprising:

9. The megakaryocyte or its precursor contains a nucleic acid encoding a chimeric platelet receptor, and the chimeric platelet receptor is i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, The manipulated megakaryocyte or precursor thereof according to any one of claims 1 to 8, comprising:

10. Engineered platelets produced by engineered megakaryocytes or their progenitor cells according to any one of claims 1 to 9.

11. The manipulated platelets have reduced thrombus-forming ability. a) A chimeric platelet receptor, wherein the chimeric platelet receptor is as follows: i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, Chimeric platelet receptors including; b) A nucleic acid encoding a chimeric platelet receptor, wherein the chimeric platelet receptor is as follows: i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, Nucleic acids encoding chimeric platelet receptors; c) A vector comprising a nucleic acid encoding a chimeric platelet receptor, wherein the chimeric platelet receptor is as follows: i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, A vector containing nucleic acids encoding a chimeric platelet receptor; and / or d) A viral vector or viral particle comprising a nucleic acid encoding a chimeric platelet receptor, wherein the chimeric platelet receptor is: i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, Viral vectors or viral particles containing nucleic acids encoding chimeric platelet receptors. The manipulated platelet according to claim 10, comprising:

12. An engineered megakaryocyte or its progenitor cell according to any one of claims 1 to 9, further comprising cargo for targeted delivery, or an engineered platelet according to claim 10 or 11.

13. The aforementioned cargo, i) Below: a) Proteins or peptides; b) Therapeutic agents; c) toxin; d) Nucleic acids, optionally therapeutic RNA or mRNA; e) Cosmetics; f) Soluble cargo; g) Membrane-bound cargo; h) Contrast agent; i) CRISPR / CAS system; j) Guide RNA; k) Small molecules, or small molecule drugs as an option; l) A passively loaded therapeutic agent; m) Genetically encoded therapeutic agents; o) Passively loaded contrast agents; and / or p) Genetically coded contrast agents is and / or ii) Below: q) Including α-granule localization signals, The manipulated megakaryocyte or its progenitor cell, or manipulated platelet, according to claim 12.

14. A delivery system comprising an engineered megakaryocyte or its progenitor cell according to any one of claims 1 to 13, or an engineered platelet, and a cargo, wherein the cargo is i) Below: a) Proteins or peptides; b) Therapeutic agents; c) toxin; d) Nucleic acids, optionally therapeutic RNA or mRNA; e) Cosmetics; f) Soluble cargo; g) Membrane-bound cargo; h) Contrast agent; i) CRISPR / CAS system; j) Guide RNA; k) Small molecules, or small molecule drugs as an option; l) A passively loaded therapeutic agent; m) Genetically encoded therapeutic agents; o) Passively loaded contrast agents; and / or p) Genetically coded contrast agents is and / or ii) Below: q) Including α-granule localization signals, Delivery system.

15. A delivery system comprising an engineered megakaryocyte or its progenitor cell, or an engineered platelet, according to any one of claims 1 to 13, and a cargo, wherein the engineered megakaryocyte or its progenitor cell, or the engineered platelet, a) A chimeric platelet receptor, wherein the chimeric platelet receptor is as follows: i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, Chimeric platelet receptors including; b) A nucleic acid encoding a chimeric platelet receptor, wherein the chimeric platelet receptor is as follows: i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, Nucleic acids encoding chimeric platelet receptors; c) A vector comprising a nucleic acid encoding a chimeric platelet receptor, wherein the chimeric platelet receptor is as follows: i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, A vector containing nucleic acids encoding a chimeric platelet receptor; and / or d) A viral vector or viral particle comprising a nucleic acid encoding a chimeric platelet receptor, wherein the chimeric platelet receptor is: i) A platelet-stimulating domain, comprising an intracellular domain including a domain from an immune receptor tyrosine-based activation motif (ITAM) receptor, ii) A heterologous targeting domain that recognizes and binds to a target, Viral vectors or viral particles containing nucleic acids encoding chimeric platelet receptors. This includes, and the cargo is optional, i) Below: a) Proteins or peptides; b) Therapeutic agents; c) toxin; d) Nucleic acids, optionally therapeutic RNA or mRNA; e) Cosmetics; f) Soluble cargo; g) Membrane-bound cargo; h) Contrast agent; i) CRISPR / CAS system; j) Guide RNA; k) Small molecules, or small molecule drugs as an option; l) A passively loaded therapeutic agent; m) Genetically encoded therapeutic agents; o) Passively loaded contrast agents; and / or p) Genetically coded contrast agents is and / or ii) Below: q) Including α-granule localization signals, Delivery system.

16. A therapeutic delivery system comprising an engineered megakaryocyte or its progenitor cell according to any one of claims 1 to 13, or an engineered platelet, and a cargo, wherein the cargo is a therapeutic agent.

17. The aforementioned cargo, a) Packaged within α-granules; b) Includes α-granule localization signals; c) Coating the surface of megakaryocytes or platelets; d) It is a protein; e) It is a nucleic acid; and / or f) Small molecules that optionally coat the surface, The therapeutic delivery system according to claim 16.

18. A method for producing manipulated platelets according to any one of claims 10 to 13, comprising incubating manipulated megakaryocytes or their precursor cells according to any one of claims 1 to 13 under conditions that induce platelet production, and optionally further comprising loading the platelets with the therapeutic agent by incubating the platelets with the therapeutic agent.

19. Manipulated megakaryocytes or their progenitor cells, and / or manipulated platelets, according to any one of claims 1 to 13, for use in medicine.

20. Manipulated megakaryocytes or their progenitor cells, and / or manipulated platelets, according to any one of claims 1 to 13, for use in the delivery of therapeutic cargo or imaging cargo, or in the treatment or prevention of cancer, autoimmune diseases, and / or infections.

21. below: The manipulated megakaryocyte according to any one of claims 1 to 13, The manipulated platelets according to any one of claims 10 to 13, and / or Therapeutic agents and / or contrast agents, A kit containing two or more of the following.

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