WO2020113063A1 - Single domain antibodies against cll-1 - Google Patents
Single domain antibodies against cll-1 Download PDFInfo
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- WO2020113063A1 WO2020113063A1 PCT/US2019/063691 US2019063691W WO2020113063A1 WO 2020113063 A1 WO2020113063 A1 WO 2020113063A1 US 2019063691 W US2019063691 W US 2019063691W WO 2020113063 A1 WO2020113063 A1 WO 2020113063A1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2851—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the lectin superfamily, e.g. CD23, CD72
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
- C07K2317/569—Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
- C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
- C07K2319/41—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a Myc-tag
Definitions
- CLL-1 also known as MICL, and Clecl2A
- AML acute myeloid leukemia
- CSCs cancer stem cells
- AML remains a major therapeutic challenge and an unmet need in hematologic oncology.
- AML is a disease resulting in uncontrollable accumulation of immature myeloid blasts in the bone marrow and peripheral blood, and the disease has multiple subtypes that contribute to the challenge in developing an encompassing targeted therapy.
- novel therapies approved for AML there remains a need for novel therapeutics for AML, such as therapeutics that target CLL-1.
- the present disclosure provides, among other things, antibodies, or antigen binding fragments thereof, that bind (e.g., selectively bind) CLL-1, compositions useful for binding CLL-1, and methods for treating disease comprising administration of such antibodies, or antigen binding fragments thereof.
- the present disclosure provides an antibody, or antigen binding fragment thereof, comprising a VHH described herein.
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having the amino acid sequence of any one of SEQ ID Nos:3-25, or a fragment thereof.
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of the C-terminal amino acids
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3- 25, wherein the portion lacks all of the C-terminal amino acids
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3- 25, wherein the portion lacks one or more of the C-terminal amino acids
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3- 25, wherein the portion lacks all of the C-terminal amino acids
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of the C-terminal amino acids
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks all of the C-terminal amino acids
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of the C-terminal amino acids
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks all of the C-terminal amino acids
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH
- the present disclosure provides an antibody, or antigen binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH comprising at least one CDR that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%,
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH comprising CDR1, CDR2, and/or CDR3 of any one of Groups 1-13 depicted in Table 1A and/or Table IB.
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH comprising (i) CDR1 and CDR2; (ii) CDR2 and CDR3; (iii) CDR1 and CDR3; or (iv) CDR1, CDR2, and CDR3 of any one of Groups 1-13 depicted in Table 1 A and/or Table IB.
- a VHH comprising (i) CDR1 and CDR2; (ii) CDR2 and CDR3; (iii) CDR1 and CDR3; or (iv) CDR1, CDR2, and CDR3 of any one of Groups 1-13 depicted in Table 1 A and/or Table IB.
- the present disclosure provides an antibody, or antigen-binding fragment thereof, that binds (e.g., selectively binds) CLL-1, comprising or consisting of a VHH comprising CDR1, CDR2, and CDR3 of Group 1; CDR1, CDR2, and CDR3 of Group 2; CDR1, CDR2, and CDR3 of Group 3; CDR1, CDR2, and CDR3 of Group 4; CDR1, CDR2, and CDR3 of Group 5; CDR1, CDR2, and CDR3 of Group 6; CDR1, CDR2, and CDR3 of Group 7; CDR1, CDR2, and CDR3 of Group 8; CDR1, CDR2, and CDR3 of Group 9; CDR1, CDR2, and CDR3 of Group 10; CDR1, CDR2, and CDR3 of Group 11; CDR1, CDR2, and CDR3 of Group 12; or CDR1, CDR2, and CDR3 of Group 13 depicted in Table 1A and/or Table IB.
- a VHH comprising CDR
- the present disclosure provides an antibody or antigen binding fragment thereof that binds (e.g., selectively binds) to CLL-1 at, e.g., an epitope consisting of or comprising the last 50, 40, or 30 C-terminal amino acids of SEQ ID NO: 28.
- the present disclosure provides an antibody or antigen-binding fragment thereof that binds (e.g., selectively binds) to CLL-1 at, e.g., an epitope consisting of or comprising amino acids 243 to 275 of SEQ ID NO: 28.
- the present disclosure provides an antibody or antigen-binding fragment thereof that binds (e.g., selectively binds) to CLL-1 at, e.g., an epitope consisting of or comprising amino acids 248 to 262 of SEQ ID NO: 28.
- the present disclosure provides an antibody or antigen binding fragment thereof that binds (e.g., selectively binds) to CLL-1 at, e.g., an epitope consisting of or comprising amino acids 251 to 260 of SEQ ID NO: 28.
- the present disclosure provides an antibody or antigen-binding fragment thereof that binds (e.g., selectively binds) to CLL-1 and does not compete for binding with SC02-357 antibody (e.g., consisting or comprising SEQ ID NO:26).
- the present disclosure provides an antibody or antigen-binding fragment thereof that binds (e.g., selectively binds) to CLL-1 and competes for binding to CLL-1 with an antibody or antigen binding fragment described herein (e.g., comprising or consisting of a sequence selected from the group consisting of SEQ ID NO:
- the present disclosure provides a nucleic acid sequence encoding an antibody or antigen-binding fragment thereof described herein.
- the present disclosure provides a vector comprising a nucleic acid sequence encoding an antibody or antigen-binding fragment thereof described herein.
- the present disclosure provides a host cell comprising a nucleic acid sequence encoding an antibody or antigen-binding fragment thereof described herein.
- the present disclosure provides a host cell comprising a vector comprising a nucleic acid sequence encoding an antibody or antigen-binding fragment thereof described herein.
- the present disclosure provides a method of producing an antibody, or antigen-binding fragment thereof, comprising culturing a host cell, e.g., a host cell comprising a nucleic acid encoding an antibody or antigen-binding fragment thereof described herein, under conditions suitable for expression of an antibody or antigen-binding fragment thereof.
- the present disclosure provides a method of treating a CLL-1 associated disease or disorder, the method comprising administering to a subject in need thereof an effective amount of an antibody, or antigen-binding fragment thereof, described herein, e.g., administering a composition (e.g., a pharmaceutical composition) comprising an effective amount of an antibody, or antigen-binding fragment thereof, described herein.
- affinity refers to the characteristics of a binding interaction between an antigen binding moiety (e.g., a single domain antibody described herein) and an antigen target (e.g., CLL-1) and that indicates the strength of the binding interaction.
- the measure of affinity is expressed as a dissociation constant (KD).
- an antigen binding moiety has a high affinity for an antigen target (e.g., a KD of less than about 10 7 M, less than about 10 8 M, or less than about 10 9 M).
- the term“approximately” or“about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value.
- the term“approximately” or“about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).
- an antibody refers to a polypeptide that includes at least one immunoglobulin variable region, e.g., an amino acid sequence that provides an immunoglobulin variable domain or immunoglobulin variable domain sequence.
- an antibody can include a heavy (H) chain variable region (abbreviated herein as VH), and a light (L) chain variable region (abbreviated herein as VL).
- VH heavy chain variable region
- L light chain variable region
- an antibody includes two heavy (H) chain variable regions and two light (L) chain variable regions.
- antibody encompasses antigen-binding domains or fragments of antibodies (e.g., single domain antibodies, single chain antibodies, Fab, F(ab')2, Fd, Fv, dAb fragments) as well as complete antibodies, e.g., intact immunoglobulins of types IgA, IgG, IgE, IgD, IgM (as well as subtypes thereof).
- the light chains of the immunoglobulin can be of types kappa or lambda.
- Antigen target is any molecule specifically bound by an antigen binding moiety of an antibody described herein.
- an antigen target is CLL-1.
- Constant region refers to a polypeptide that corresponds to, or is derived from, one or more constant region immunoglobulin domains of an antibody.
- a constant region can include any or all of the following
- immunoglobulin domains a CHI domain, a hinge region, a CH2 domain, a CH3 domain (derived from an IgA, IgD, IgG, IgE, or IgM), and a CH4 domain (derived from an IgE or IgM).
- Fc region refers to a dimer of two“Fc polypeptides”, each“Fc polypeptide” comprising the constant region of an antibody excluding the first constant region immunoglobulin domain.
- an“Fc region” includes two Fc polypeptides linked by one or more disulfide bonds, chemical linkers, or peptide linkers.
- Fc polypeptide refers to the last two constant region immunoglobulin domains of IgA, IgD, and IgG, and the last three constant region immunoglobulin domains of IgE and IgM, and may also include part or all of the flexible hinge N-terminal to these domains.
- “Fc polypeptide” comprises immunoglobulin domains Cgamma2 (Cy2) and Cgamma3 (Oy3) and the lower part of the hinge between Cgammal (Oyl) and Oy2.
- Fc polypeptide comprises immunoglobulin domains Calpha2 (Ca2) and Calpha3 (Ca3) and the lower part of the hinge between Calphal (Cal) and Ca2.
- An Fc region can be synthetic, recombinant, or generated from natural sources such as IVIG.
- Identity refers to the overall relatedness between between nucleic acid molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptides.
- nucleic acid molecules e.g., DNA molecules and/or RNA molecules
- polypeptides e.g., DNA molecules and/or RNA molecules
- nucleic acids or polypeptides are considered to be “substantially identical” to one another if their sequences are at least 25%, 30%, 35%, 40%,
- Calculation of the percent identity of two nucleic acid or polypeptide sequences can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-identical sequences can be disregarded for comparison purposes).
- the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of a reference sequence.
- the nucleotides at corresponding positions are then compared. When a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, then the molecules are identical at that position.
- the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences.
- the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm.
- the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0).
- nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
- the percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix.
- Immunoglobulin single variable domain means an immunoglobulin variable domain which is capable of specifically binding to an epitope of the antigen without pairing with an additional variable immunoglobulin domain.
- immunoglobulin single variable domain or“single variable domain”, as used herein, means an immunoglobulin variable domain which is capable of specifically binding to an epitope of the antigen without pairing with an additional variable immunoglobulin domain.
- immunoglobulin single variable domain in the meaning of the present disclosure is“domain antibody”, such as the immunoglobulin single variable domains VH and VL (VH domains and VL domains).
- Another example of an immunoglobulin single variable domain is“VHH domain” (or simply“VHH”) from camelids, as described herein.
- Immunoglobulin variable domain means an immunoglobulin domain that is or includes four “framework regions” (referred to in the art and herein as“framework region 1” or“FR1”; as “framework region 2” or“FR2”; as“framework region 3” or“FR3”; and as“framework region 4” or“FR4”, respectively); which framework regions are interrupted by three“complementarity determining regions” or“CDRs” (referred to in the art and herein as“complementarity determining region 1” or“CDR1”; as“complementarity determining region 2” or“CDR2”; and as“complementarity determining region 3” or“CDR3”, respectively).
- the general structure or sequence of an immunoglobulin variable domain can be indicated as follows: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4.
- Ka refers to an association rate of a particular antigen binding moiety and an antigen target to form an antigen binding moiety/antigen target complex.
- Kd refers to a dissociation rate of a particular antigen binding moiety/antigen target complex.
- KD refers to a dissociation constant, which is obtained from the ratio of Kd to K a (i.e., Kd/K a ) and is expressed as a molar concentration (M). KD values can be determined using methods well established in the art, e.g., by using surface plasmon resonance, or using a biosensor system such as a Biacore® system.
- “specific binding”, or“specifically binds” refers, with respect to an antigen binding moiety and an antigen target, preferential association of an antigen binding moiety to an antigen target and not to an entity that is not the antigen target. A certain degree of non-specific binding may occur between an antigen binding moiety and a non-target.
- an antigen binding moiety selectively binds an antigen target if binding between the antigen binding moiety and the antigen target is greater than 2-fold, greater than 5-fold, greater than 10-fold, or greater than 100- fold as compared with binding of the antigen binding moiety and a non-target.
- an antigen binding moiety selectively binds an antigen target if the binding affinity is less than about 10 5 M, less than about 10 6 M, less than about 10 7 M, less than about 10 8 M, or less than about 10 9 M.
- Subject means any subject for whom diagnosis, prognosis, or therapy is desired.
- a subject can be a mammal, e.g. , a human or non-human primate (such as an ape, monkey, orangutan, or chimpanzee), a dog, cat, guinea pig, rabbit, rat, mouse, horse, cattle, or cow.
- a human or non-human primate such as an ape, monkey, orangutan, or chimpanzee
- a dog cat, guinea pig, rabbit, rat, mouse, horse, cattle, or cow.
- therapeutically effective amount refers to an amount of an antibody or composition described herein that confers a therapeutic effect on a treated subject, at a reasonable benefit/risk ratio applicable to any medical treatment.
- the therapeutic effect may be objective (i.e., measurable by some test or marker) or subjective (i.e., subject gives an indication of or feels an effect).
- therapeutically effective amount refers to an amount of an antibody or composition effective to treat, ameliorate, or prevent a particular disease or condition, or to exhibit a detectable therapeutic or preventative effect, such as by ameliorating symptoms associated with the disease, preventing or delaying the onset of the disease, and/or also lessening the severity or frequency of symptoms of the disease.
- a therapeutically effective amount can be administered in a dosing regimen that may comprise multiple unit doses.
- a therapeutically effective amount (and/or an appropriate unit dose within an effective dosing regimen) may vary, for example, depending on route of administration, on combination with other pharmaceutical agents.
- the specific therapeutically effective amount (and/or unit dose) for any particular subject may depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific pharmaceutical agent employed; the specific composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and/or rate of excretion or metabolism of the specific therapeutic molecule employed; the duration of the treatment; and like factors as is well known in the medical arts.
- treatment refers to any administration of an antibody or composition described herein that partially or completely alleviates, ameliorates, relieves, inhibits, delays onset of, reduces severity of and/or reduces incidence of one or more symptoms or features of a particular disease, disorder, and/or condition.
- Such treatment may be of a subject who does not exhibit signs of the relevant disease, disorder and/or condition and/or of a subject who exhibits only early signs of the disease, disorder, and/or condition.
- such treatment may be of a subject who exhibits one or more established signs of the relevant disease, disorder and/or condition.
- Figure 1 shows binding of certain VHH clones to CLEC12A.
- Figures 2A-2C show U937 cell binding by certain VHH clones.
- Figures 3A-3C show determination of EC50s of certain VHH clones.
- Figures 4A and 4B show E1937 cell binding by certain VHH clones.
- Figures 5A-5D show E1937 cell binding by certain VHH clones.
- Figures 6A-6C show E1937 cell binding by certain VHH clones.
- Figures 7A-7D shows determination of distinct or similar epitopes among certain
- VHH clones VHH clones
- Figure 8 demonstrates VHH clone 1H1 binds a linear epitope of Clecl2A.
- Figure 9 demonstrates epitope differences between an anti-Clecl2A scFv and certain VHH clones.
- Figures 10A and 10B demonstrate the ability of VHH clone 2H3 to recognize recombinant Clecl2A proteins.
- Figures 11 A and 1 IB demonstrate mass spectrometry identified epitopes of an anti-Clecl2A scFv and VHH clone 2H3 (Note: amino acid numbering in figure based on only extracellular domain amino acids (i.e., the 200 amino acids 65-25 of the full length sequence).
- Figure 12 shows VHH clone 2H3 binds specifically to Clecl2A.
- the present disclosure is based, in part, on the discovery of single domain antibodies that selectively bind to CLL-1/CLEC12A.
- the disclosure also relates to nucleic acids encoding said antibodies, or antigen-binding fragments thereof, cells comprising such nucleic acids, and methods of use.
- Single domain antibodies are antibodies whose complementary determining regions are part of a single domain polypeptide. Examples include, but are not limited to, heavy chain antibodies, antibodies naturally devoid of light chains, single domain antibodies derived from conventional 4-chain antibodies, engineered antibodies and single domain scaffolds other than those derived from antibodies. Single domain antibodies may be any of the art known, or any future single domain antibodies. Single domain antibodies may be derived from any species including, but not limited to mouse, human, camel, llama, goat, rabbit, bovine. According to one aspect of the disclosure, a single domain antibody as used herein is a naturally occurring single domain antibody known as heavy chain antibody devoid of light chains. Such single domain antibodies are disclosed in, e.g., WO 94/04678.
- variable domains derived from a heavy chain antibody naturally devoid of light chain is referred to herein as a“VHH” or“nanobody”.
- VHH can be derived from antibodies raised in Camelidae species, for example in camel, dromedary, llama, vicuna, alpaca and guanaco.
- Other species besides Camelidae may produce heavy chain antibodies naturally devoid of light chain; such VHHs are within the scope of the disclosure.
- VHH domains from Camelids are numbered according to the general numbering for VH domains given by Kabat et ah,“Sequence of proteins of immunological interest”, US Public Health Services, NIH (Bethesda, MD), Publication No 91-3242 (1991); see also Riechmann et ah, J. Immunol. Methods 231 :25-38 (1999).
- FR1 comprises the amino acid residues at positions 1-30
- CDR1 comprises the amino acid residues at positions 31-35
- FR2 comprises the amino acids at positions 36-49
- CDR2 comprises the amino acid residues at positions 50-65
- FR3 comprises the amino acid residues at positions 66-94
- CDR3 comprises the amino acid residues at positions 95-102
- FR4 comprises the amino acid residues at positions 103-113.
- VHH domains that the total number of amino acid residues in each of the CDRs may vary and may not correspond to the total number of amino acid residues indicated by the Kabat numbering (that is, one or more positions according to the Kabat numbering may not be occupied in the actual sequence, or the actual sequence may contain more amino acid residues than the number allowed for by the Kabat numbering).
- the numbering according to Kabat may or may not correspond to the actual numbering of the amino acid residues in the actual sequence.
- Alternative methods for numbering the amino acid residues of VH domains which methods can also be applied in an analogous manner to VHH domains, are known in the art. However, in the present disclosure, claims and figures, the numbering according to Kabat and applied to VHH domains as described above will be followed, unless indicated otherwise.
- the disclosure provides a CLL-1 binding antibody that is or includes a VHH having the amino acid sequence of any one of SEQ ID Nos:3-25, or a fragment thereof (e.g., a CLL-1 binding fragment thereof).
- each of SEQ ID Nos:3-25 includes VHH amino acids at the N- terminus, and the following amino acids at the C-terminus: (i) a linker of 9 amino acids
- TSGPGGQGA a myc-tag
- EQKLISEEDL a linker of 2 amino acids
- GA a hexa-histidine tag
- GAS an additional 3 amino acids
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of (i)-(v) (and/or lacks a portion of one or more of (i)-(v)).
- a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of (i)-(v) (and/or lacks a portion of one or more of (i)-(v)).
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of the C-terminal amino acids
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL- 1, that is or includes a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks all of the C-terminal amino acids T S GPGGQGAEQKLI SEEDLGAHHHHHHGA S depicted in each of SEQ ID Nos:3-25.
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of (i)-(v) (and/or lacks a portion of one or more of (i)-(v)), and wherein the portion lacks one or more (e.g., 1, 2, 3, 4, 5, or more), additional amino acids (i.e., other than an amino acid included in (i)-(v)).
- a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of (i)-(v) (and/or lacks a portion of one or more of (i)-(v)), and wherein the portion lacks one or more (e.g., 1, 2, 3,
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of the C-terminal amino acids
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL- 1, that is or includes a VHH having a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks all of the C-terminal amino acids T S GPGGQGAEQKLI SEEDLGAHHHHHHGA S depicted in each of SEQ ID Nos:3-25, and wherein the portion lacks one or more (e.g., 1, 2, 3, 4, 5, or more), additional amino acids.
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3- 25, wherein the portion lacks one or more of (i)-(v) (and/or lacks a portion of one or more of (i)- (v)).
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having an amino acid sequence that is at least 85%,
- a portion e.g., a CLL-1 binding portion of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of the C-terminal amino acids
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL- 1, that is or includes a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks all of the C-terminal amino acids T SGPGGQGAEQKLISEEDLGAHHHHGAS depicted in each of SEQ ID Nos:3-25.
- a portion e.g., a CLL-1 binding portion
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3- 25, wherein the portion lacks one or more of (i)-(v) (and/or lacks a portion of one or more of (i)- (v)), and wherein the portion lacks one or more (e.g., 1, 2, 3, 4, 5, or more), additional amino acids (i.e., other than an amino acid included in (i)-(v)).
- a portion e.g., a CLL-1 binding portion of the amino acid sequence of any one of SEQ ID Nos:3- 25, wherein the portion lacks one or more of (i)-(v) (
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of the C-terminal amino acids T S GPGGQGAEQKLI SEEDLGAHHHHHHGA S depicted in each of SEQ ID Nos:3-25, and wherein the portion lacks one or more (e.g., 1, 2, 3, 4, 5, or more), additional amino acids.
- a portion e.g., a CLL-1 binding portion of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks one or more of the C-terminal amino acids T S GPGGQGAEQKLI SEED
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH having an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion (e.g., a CLL-1 binding portion) of the amino acid sequence of any one of SEQ ID Nos:3-25, wherein the portion lacks all of the C-terminal amino acids TSGPGGQGAEQKLISEEDLGAHHHHGAS depicted in each of SEQ ID Nos:3-25, and wherein the portion lacks one or more (e.g., 1, 2, 3, 4, 5, or more), additional amino acids.
- a portion e.g., a CLL-1 binding portion
- the portion lacks all of the C-terminal amino acids TSGPGGQGAEQKLISEEDLGAHHHHHHGAS depicted in each of SEQ ID Nos:3-25, and wherein the portion
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH comprising at least one CDR (e.g., CDR1, CDR2, and/or CDR3) depicted in any one of SEQ ID Nos:3-25.
- a VHH comprising at least one CDR (e.g., CDR1, CDR2, and/or CDR3) depicted in any one of SEQ ID Nos:3-25.
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH comprising a portion of at least one CDR (e.g., CDR1, CDR2, and/or CDR3) depicted in any one of SEQ ID Nos:3-25, wherein the portion lacks 1, 2, 3, 4, 5, or more amino acids of a CDR depicted in any one of SEQ ID Nos:3-25.
- a VHH comprising a portion of at least one CDR (e.g., CDR1, CDR2, and/or CDR3) depicted in any one of SEQ ID Nos:3-25, wherein the portion lacks 1, 2, 3, 4, 5, or more amino acids of a CDR depicted in any one of SEQ ID Nos:3-25.
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH comprising at least one CDR that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a CDR (e.g., CDR1, CDR2, and/or CDR3) depicted in any one of SEQ ID Nos:3-25.
- a CDR e.g., CDR1, CDR2, and/or CDR3
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH comprising an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion of at least one CDR (e.g., CDR1, CDR2, and/or CDR3) depicted in any one of SEQ ID Nos:3-25, wherein the portion lacks 1, 2, 3, 4, 5, or more amino acids of a CDR depicted in any one of SEQ ID Nos: 3 -25.
- a VHH comprising an amino acid sequence that is at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a portion of at least one CDR (e.g., CDR1, CDR2, and/or CDR3) depicted in any one of SEQ ID Nos:3-25, wherein
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH comprising CDR1, CDR2, and/or CDR3 of any one of Groups 1-13 depicted in Table 1A and/or Table IB.
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH comprising (i) CDR1 and CDR2; (ii) CDR2 and CDR3; (iii) CDR1 and CDR3; or (iv) CDR1, CDR2, and CDR3 of any one of Groups 1-13 depicted in Table 1A and/or Table IB (e.g., wherein the CDRs are from one particular Group, or wherein the CDRs are selected from two or more different Groups).
- a VHH comprising (i) CDR1 and CDR2; (ii) CDR2 and CDR3; (iii) CDR1 and CDR3; or (iv) CDR1, CDR2, and CDR3 of any one of Groups 1-13 depicted in Table 1A and/or Table IB (e.g., wherein the CDRs are from one particular Group, or wherein the CDRs are selected from two or more different
- the disclosure provides an antibody that binds (e.g., selectively binds) CLL-1, that is or includes a VHH comprising CDR1, CDR2, and CDR3 of Group 1; CDR1, CDR2, and CDR3 of Group 2; CDR1, CDR2, and CDR3 of Group 3; CDR1, CDR2, and CDR3 of Group 4; CDR1, CDR2, and CDR3 of Group 5; CDR1, CDR2, and CDR3 of Group 6; CDR1, CDR2, and CDR3 of Group 7; CDR1, CDR2, and CDR3 of Group 8; CDR1, CDR2, and CDR3 of Group 9; CDR1, CDR2, and CDR3 of Group 10; CDR1, CDR2, and CDR3 of Group 11; CDR1, CDR2, and CDR3 of Group 12; or CDR1, CDR2, and CDR3 of Group 13, as depicted in Table 1 A and/or Table IB.
- VHH comprising CDR1, CDR2, and CDR3 of Group 1; CDR
- Table IB CDRs as identified based on IMGT numbering and ANARCI software
- the present disclosure provides an antibody or antigen binding fragment thereof that binds (e.g., selectively binds) to CLL-1 at, e.g., an epitope in the last 50, 40, or 30 amino acids of SEQ ID NO: 28.
- the present disclosure provides an antibody or antigen-binding fragment thereof that binds (e.g., selectively binds) to CLL-1 at, e.g., an epitope within amino acids 243 to 275 of SEQ ID NO: 28.
- the present disclosure provides an antibody or antigen-binding fragment thereof that binds (e.g., selectively binds) to CLL-1 at, e.g., an epitope within amino acids 248 to 262 of SEQ ID NO: 28. In some embodiments, the present disclosure provides an antibody or antigen binding fragment thereof that binds (e.g., selectively binds) to CLL-1 at, e.g., an epitope within amino acids 251 to 260 of SEQ ID NO: 28. In some embodiments, the present disclosure provides an antibody or antigen-binding fragment thereof that binds (e.g., selectively binds) to CLL-1 and does not compete for binding with SC02-357.
- the present disclosure provides an antibody or antigen-binding fragment thereof that binds (e.g., selectively binds) to CLL-1 and that competes for binding to CLL-1 with an antibody or antigen binding fragment described herein (e.g., comprising a sequence selected from the group consisting of SEQ ID NO: 3 to SEQ ID NO: 25).
- any such CDR sequence may be readily combined, e.g., using molecular biology techniques, with any other polypeptide (e.g., antibody) sequences or domains provided herein or otherwise known in the art, including any framework regions, CDRs, or constant domains, or portions thereof as disclosed herein or otherwise known in the art, as may be present in an antibody or binding molecule of any format as disclosed herein or otherwise known in the art.
- polypeptide e.g., antibody sequences or domains provided herein or otherwise known in the art, including any framework regions, CDRs, or constant domains, or portions thereof as disclosed herein or otherwise known in the art, as may be present in an antibody or binding molecule of any format as disclosed herein or otherwise known in the art.
- Antibodies or fragments can be produced by any method known in the art for synthesizing antibodies (see, e.g., Harlow et ak, Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Brinkman et ak, 1995, J. Immunol. Methods 182:41-50; WO 92/22324; WO 98/46645).
- Chimeric antibodies can be produced using methods described in, e.g., Morrison, 1985, Science 229:1202, and humanized antibodies by methods described in, e.g., U.S. Pat. No. 6,180,370.
- compositions and methods described herein are bispecific antibodies and multivalent antibodies, as described in, e.g., Segal et ak, J. Immunol. Methods 248: 1-6 (2001); and Tutt et ak, J. Immunol. 147: 60 (1991).
- the disclosure provides fusion proteins comprising (i) one or more single domain antibodies, or antigen-binding fragments thereof, described herein (e.g., one or more CDRs described herein), and (ii) one or more additional polypeptides.
- a fusion protein can include one or more single domain antibodies described herein and a constant region or Fc region described herein.
- one or more single domain antibodies, or antigen-binding fragments thereof, described herein can be conjugated noncovalently or covalently, e.g., fused, to an antigen (e.g., an antigen target for a cellular therapeutic, e.g., a CAR-T cell or antibody drug conjugate) as described in, e.g., WO2017/075537, WO2017/075533, WO2018156802, and WO2018156791.
- an antigen e.g., an antigen target for a cellular therapeutic, e.g., a CAR-T cell or antibody drug conjugate
- the disclosure provides a fusion protein comprising one or more VHH as described herein and one or more additional polypeptides.
- an additional polypeptide comprises an additional antibody or fragment thereof. Additional antibodies include, e.g., intact IgG, IgE and IgM, bi- or multi- specific antibodies (e.g.,
- KALBITOR® Exemplary additional antibodies are listed in Table 4.
- an additional antibody targets PD-1, TIM-3, LAG-3, IDO, A2AR, TGFbeta, CD47, or another protein involved in an immunosuppressive pathway.
- an additional polypeptide comprises or consists of all or a portion of a tumor associated antigen (TAA) or tumor specific antigen (TSA).
- TSA or TAA antigens include differentiation antigens such as MART-l/MelanA (MART-I), gplOO (Pmel 17), tyrosinase, TRP-1, TRP-2 and tumor-specific multilineage antigens such as MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, pi 5; overexpressed embryonic antigens such as CEA; overexpressed oncogenes and mutated tumor-suppressor genes such as p53, Ras, HER-2/neu; unique tumor antigens resulting from chromosomal translocations such as BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR; and viral antigens, such as the Epstein Barr virus antigens EBVA and the
- tumor antigens include TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, erbB, pl85erbB2, pl80erbB-3, c-met, nm-23Hl, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1,
- an additional polypeptide comprises or consists of all or a portion of a tumor antigen selected from CD 19, CD20, CD22, CD30, CD72, CD 180, CD171 (LI CAM), CD123, CD133, CD138, CD37, CD70, CD79a, CD79b, CD56, CD74, CD166,
- CD71 CD71, CLL-1/CLECK12A, ROR1, Glypican 3 (GPC3), Mesothelin, CD33/IL3Ra, c-Met,
- an additional polypeptide comprises or consists of all or a portion of a B cell specific marker selected from CD 19, CD20, CD21, CD22, CD23, CD24, CD40, CD72, CD 180, ROR1, BCMA, CD79a, and CD79b (see, e.g., LeBien et al., Blood 112: 1570-1580 (2008)).
- the disclosure provides antibodies conjugated to a therapeutic moiety, such as a cytotoxin, a biologically active protein (e.g., one or more peptide or one or more cytokine) or a radioisotoperadiotoxin.
- a therapeutic moiety such as a cytotoxin, a biologically active protein (e.g., one or more peptide or one or more cytokine) or a radioisotoperadiotoxin.
- a therapeutic moiety such as a cytotoxin, a biologically active protein (e.g., one or more peptide or one or more cytokine) or a radioisotoperadiotoxin.
- immunococonjugates Immunoconjugates that include one or more cytotoxins are referred to as“immunotoxins”.
- a cytotoxin or cytotoxic agent includes any agent that is detrimental to (e.g., kills) cells.
- Examples include taxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1- dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof.
- Therapeutic agents that can be conjugated also include, for example,
- antimetabolites e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine
- alkylating agents e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU)
- cyclothosphamide busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin
- anthracyclines e.g., daunorubicin (formerly daunomycin) and doxorubicin
- antibiotics e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)
- anti-mitotic agents e.g., vincristine and
- therapeutic cytotoxins that can be conjugated to antibodies described herein include duocarmycins, calicheamicins, maytansines and auristatins, and derivatives thereof.
- Cytotoxins can be conjugated to antibodies using linker technology available in the art.
- linker types that have been used to conjugate a cytotoxin to antibodies include, but are not limited to, hydrazones, thioethers, esters, disulfides and peptide-containing linkers.
- a linker can be chosen that is, for example, susceptible to cleavage by low pH within the lysosomal compartment or susceptible to cleavage by proteases, such as proteases preferentially expressed in tumor tissue such as cathepsins (e.g., cathepsins B, C, D).
- Antibodies described herein can also be conjugated to a radioactive isotope to generate cytotoxic radiopharmaceuticals, also referred to as radioimmunoconjugates.
- radioactive isotopes that can be conjugated to antibodies for use diagnostically or
- radioimmunoconjugates include, but are not limited to, iodine 131 , indium 111 , yttrium 90 and lutetium 177 .
- Methods for preparing radioimmunoconjugates are established in the art. Examples of radioimmunoconjugates are commercially available, including ZevalinTM (Spectrum
- BexxarTM (formally sold by GlaxoSmithKline), and similar methods can be used to prepare radioimmunoconjugates using antibodies described herein.
- the present disclosure includes nucleotide sequences encoding one or more antibodies described herein (e.g., a VHH described herein), or portion thereof (e.g., one or more CDRs described herein), and/or one or more fusion proteins described herein.
- nucleotide sequences may be present in a vector.
- nucleotides may be present in the genome of a cell, e.g., a cell of a subject in need of treatment or a cell for production of an antibody, e.g. a mammalian cell for production of an antibody.
- CLL-1 Human C-type lectin-like molecule-1 (CLL-1), also known as MICL or
- CLEC12A is a type II transmembrane glycoprotein and member of the large family of C-type lectin-like receptors involved in immune regulation.
- CLL-1 has previously been identified from myeloid-derived cells.
- the intracellular domain of CLL-1 contains an immunotyrosine-based inhibition motif (ITIM) and a YXXM motif. Phosphorylation of ITIM-containing receptors on a variety of cells results in inhibition of activation pathways through recruitment of protein tyrosine phosphatases SHP-1, SHP-2 and SHIP.
- the YXXM motif has a potential SH2 domain binding site for the p85 subunit of PI-3 kinase, 13 which has been implicated in cellular activation pathways, revealing a potential dual role of CLL-1 as an inhibitory and activating molecule on myeloid cells. Indeed, association of CLL-1 with SHP-1 and SHP-2 has been demonstrated experimentally in transfected and myeloid-derived cell lines.
- CLL-1 is also present on the majority of leukemic stem cells in the CD34+/CD38- compartment in AML but absent from CD34+/CD38- cells in normal and in regenerating bone marrow controls, which aids the discrimination between normal and leukemic stem cells.
- CLL-1 Associated Disorders The nucleotide and protein sequences of CLL-1 are known for many species.
- the human sequences can be found at Genbank accession number AF247788.1 (coding sequence shown in SEQ ID NO:l) and Uniprot accession number Q5QGZ9 (SEQ ID NO:2).
- the extracellular domain comprises approximately amino acids 65-265
- the transmembrane domain comprises approximately amino acids 44-64
- the cytoplasmic domain comprises approximately amino acids 1-43.
- the stalk domain of human CLL-1 spans amino acids 65-139
- the C lectin domain spans amino acids 140-249, both with reference to the sequence shown in SEQ ID NO:2.
- CLL-1 variants e.g., species homologs, allelic variants, etc.
- CLL-1 Associated Disorders can be optimally aligned, e.g., for identification of conserved residues and domains.
- the antibodies and/or fusion proteins of the disclosure can be used, e.g., to detect and/or treat CLL-1 associated disorders, i.e., diseases correlated with elevated or reduced cell surface expression of CLL-1 as compared to CLL-1 expression in a standard control (e.g., a normal, non-disease, non-cancer cell).
- CLL-1 expression is normally limited to myeloid lineage cells, e.g., dendritic cells, granulocytes, and monocytes in the peripheral blood and spleen.
- elevated CLL-1 levels are associated with cancer, in particular, in hematopoietic CSCs (e.g., LSCs), and in myeloproliferative disorders, including leukemias such as AML (acute myelogenous or myeloproliferative leukemia), MDS (myelodysplastic syndrome), myelofibrosis, CMML (chronic myelomonocytic leukemia), multiple myeloma, plasmacytoma, and CML (chronic myelogenous or myeloproliferative leukemia).
- leukemias such as AML (acute myelogenous or myeloproliferative leukemia), MDS (myelodysplastic syndrome), myelofibrosis, CMML (chronic myelomonocytic leukemia), multiple myeloma, plasmacytoma, and CML (chronic myelogenous or myeloproliferative leukemia).
- AML cells can be characterized and distinguished from other cells by detecting cell surface marker expression. Aside from being CLL-1+, AML cells can be CD33+(though some are CD33-), CD45+, and CDw52+. AML blasts (including LSCs) are typically
- CD34+CD38- HSCs and LSCs can be characterized by expression of CD34, but the former do not express CLL-1.
- MDS cells can be characterized by expression of CDS, CD7, CD13, and CD34.
- CML cells can be characterized by expression of 7-ADD, CD33, CD34, and CD38.
- MDS Myelodysplastic Syndromes
- preleukemia refractory anemia
- refractory dysmyelopoietic anemia refractory dysmyelopoietic anemia
- DMPS dysmyelopoietic syndrome
- myelodysplasia myelodysplasia
- DMPS can be characterized by presence of megablastoids, megarkaryocyte dysplasia, and an increase in number of abnormal blast cells, reflective of enhanced granulocyte maturation process.
- Patients with DMPS typically show chromosomal abonormalities similar to those found in acute myeloid leukemia and progress to acute myeloid leukemia in a certain fraction of afflicted patients.
- Chronic myeloproliferative disorders are a collection of conditions that can be characterized by increased number of mature and immature granulocytes, erythrocytes, and platelets. Chronic myeloproliferative disorders can transition to other forms within this group, with a tendency to terminate in acute myeloid leukemia. Specific diseases within this group include polycythemia vera, chronic myeloid leukemia, agnogenic myeloid leukemia, essential thrombocythemia, and chronic neutrophilic leukemia.
- Myelofibrosis can be characterized by scarring of the bone marrow that can result in reduced number of red and white blood cells, and platelets. Myelofibrotic scarring can result from leukemia, but can have other causes, such as thrombocytosis or adverse drug effects.
- an antibody and/or fusion protein described herein treats, alleviates, reduces the prevalence of, reduces the frequency of, or reduces the level or amount of one or more symptoms or biomarkers of a CLL-1 -associated disorder. Specific symptoms and progression of symptoms vary among subjects.
- an antibody and/or fusion protein described herein is administered to a subject in need thereof, e.g., a subject having a CLL-1 -associated disorder.
- administering results in a decrease in the prevalence, frequency, level, and/or amount of one or more symptoms or biomarkers of a CLL-1 -associated disorder, e.g., a decrease of at least 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100% of one or more symptoms or biomarkers as compared to a prior
- an effective dose of an antibody and/or fusion protein as described herein may be, e.g., less than 1,000 mg/dose, e.g., less than 900 mg/dose, 800 mg/dose, 700 mg/dose, 600 mg/dose, 500 mg/dose, 550 mg/dose, 400 mg/dose, 350 mg/dose, 300 mg/dose, 200 mg/dose, 100 mg/dose, 50 mg/dose, 25 mg/dose, or less.
- an antibody and/or fusion protein as described herein may be effectively or usefully administered at a frequency that is less than once per week, e.g., less than once every week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or year.
- an antibody and/or fusion protein described herein can be used in a number of diagnostic and/or therapeutic applications.
- detectably-labeled versions of antibodies as described herein can be used in assays to detect the presence or amount of CLL-1 in a sample (e.g., a biological sample).
- Antibodies and/or fusion proteins described herein can be used in in vitro assays for studying inhibition of CLL-1 activity.
- an antibody and/or fusion protein described herein can be used as a positive control in an assay designed to identify additional novel compounds that inhibit CLL-1 or otherwise are useful for treating a CLL-1 -associated disorder.
- Antibodies and/or fusion proteins described herein may be used in monitoring a subject, e.g., a subject having, suspected of having, at risk of developing, or under treatment for one or more CLL-1 -associated disorders. Monitoring may include determining the amount or activity of CLL-1 in a subject, e.g., in the serum of a subject. In some embodiments, the evaluation is performed at least one (1) hour, e.g., at least 2, 4, 6, 8, 12, 24, or 48 hours, or at least 1 day, 2 days, 4 days, 10 days, 13 days, 20 days or more, or at least 1 week, 2 weeks, 4 weeks, 10 weeks, 13 weeks, 20 weeks or more, after an administration of an antibody and/or fusion protein as described herein.
- the subject can be evaluated in one or more of the following periods: prior to beginning of treatment; during the treatment; or after one or more elements of the treatment have been administered. Evaluation can include evaluating the need for further treatment, e.g., evaluating whether a dosage, frequency of administration, or duration of treatment should be altered. It can also include evaluating the need to add or drop a selected therapeutic modality, e.g., adding or dropping any of the treatments for a CLL-1 -associated disorder described herein. Measuring Interactions of Antibodies and CLL-1
- the binding properties of an antibody described herein to CLL-1 can be measured by methods known in the art, e.g., one of the following methods: BIACORE analysis, Enzyme Linked Immunosorbent Assay (ELISA), x-ray crystallography, sequence analysis and scanning mutagenesis.
- the binding interaction of an antibody and CLL-1 can be analyzed using surface plasmon resonance (SPR).
- SPR or Biomolecular Interaction Analysis (BIA) detects bio-specific interactions in real time, without labeling any of the interactants. Changes in the mass at the binding surface (indicative of a binding event) of the BIA chip result in alterations of the refractive index of light near the surface.
- the changes in the refractivity generate a detectable signal, which are measured as an indication of real-time reactions between biological molecules.
- Methods for using SPR are described, for example, in U.S. Pat. No. 5,641,640; Raether (1988) Surface Plasmons Springer Verlag; Sjolander and Urbaniczky (1991) Anal. Chem. 63:2338- 2345; Szabo et al. (1995) Curr. Opin. Struct. Biol. 5:699-705 and on-line resources provide by BIAcore International AB (Uppsala, Sweden). Additionally, a KinExA® (Kinetic Exclusion Assay) assay, available from Sapidyne Instruments (Boise, Id.) can also be used.
- Information from SPR can be used to provide an accurate and quantitative measure of the equilibrium dissociation constant (KD), and kinetic parameters, including K 0n and Koff, for the binding of an antibody to CLL-1. Such data can be used to compare different molecules. Information from SPR can also be used to develop structure-activity relationships (SAR). Variant amino acids at given positions can be identified that correlate with particular binding parameters, e.g., high affinity.
- an antibody described herein exhibits high affinity for binding CLL-1.
- KD of an antibody as described herein for CLL-1 is less than about 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , 10 9 , lO 10 , lO 11 , 10 12 , 10 13 , 10 14 , or 10 15 M.
- KD of an antibody as described herein for CLL-1 is between 0.001 and 1 nM, e.g., 0.001 nM, 0.005 nM, 0.01 nM, 0.05 nM, 0.1 nM, 0.5 nM, or 1 nM.
- compositions e.g., pharmaceutical compositions, containing one or more antibodies and/or fusion proteins described herein, formulated together with a pharmaceutically acceptable carrier.
- a pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.
- the carrier can be suitable for intravenous, intramuscular, subcutaneous, parenteral, spinal or epidermal administration (e.g., by injection or infusion).
- the antibody may be coated in a material to protect the compound from the action of acids and other natural conditions that may inactivate the compound.
- a pharmaceutical composition may include a pharmaceutically acceptable anti oxidant.
- pharmaceutically acceptable antioxidants include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium
- metabi sulfite sodium sulfite and the like
- oil-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like
- metal chelating agents such as citric acid
- compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents.
- Pharmaceutically acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
- sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion.
- the use of such media and agents for pharmaceutically active substances is known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the pharmaceutical compositions of the invention is
- Supplementary active compounds can also be incorporated into the compositions.
- compositions typically must be sterile and stable under the conditions of manufacture and storage.
- the composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration.
- the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof.
- the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
- Sterile injectable solutions can be prepared by incorporating an antibody in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterilization microfiltration.
- dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above.
- the preferred methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
- the amount of antibody and/or fusion protein described herein that can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated, and the particular mode of administration.
- the amount of antibody and/or fusion protein that can be combined with a carrier material to produce a single dosage form will generally be that amount of the composition which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 0.01 percent to about ninety-nine percent of antibody, preferably from about 0.1 percent to about 70 percent, most preferably from about 1 percent to about 30 percent of antibody in combination with a pharmaceutically acceptable carrier.
- Dosage regimens are adjusted to provide the optimum desired response (e.g., a therapeutic response).
- Dosage unit form refers to physically discrete units suited as unitary dosages for the subjects to be treated; each unit contains a predetermined quantity of
- the dosage ranges from about 0.0001 to 100 mg/kg, and more usually 0.01 to 5 mg/kg, of the subject body weight.
- dosages can be 0.3 mg/kg body weight, 1 mg/kg body weight, 3 mg/kg body weight, 5 mg/kg body weight, 10 mg/kg body weight or 20 mg/kg body weight or within the range of 1-20 mg/kg.
- An exemplary treatment regime entails administration once per week, once every two weeks, once every three weeks, once every four weeks, once a month, once every 3 months or once every three to 6 months, or with a short administration interval at the beginning (such as once per week to once every three weeks), and then an extended interval later (such as once a month to once every three to 6 months).
- antibody and/or fusion protein can be administered as a sustained release formulation, in which case less frequent administration is required. Dosage and frequency vary depending on the half-life of the antibody and/or fusion protein in the patient. In general, human antibodies show the longest half life, followed by humanized antibodies, chimeric antibodies, and nonhuman antibodies. The dosage and frequency of administration can vary depending on whether the treatment is prophylactic or therapeutic. In prophylactic applications, a relatively low dosage is administered at relatively infrequent intervals over a long period of time. Some patients continue to receive treatment for the rest of their lives. In therapeutic applications, a relatively high dosage at relatively short intervals is sometimes required until progression of the disease is reduced or terminated, and preferably until the patient shows partial or complete amelioration of symptoms of disease. Thereafter, the patient can be administered a prophylactic regime.
- Actual dosage levels of antibody and/or fusion protein in the pharmaceutical compositions of the disclosure may be varied so as to obtain an amount of antibody and/or fusion protein that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.
- the selected dosage level will depend upon a variety of pharmacokinetic factors including the activity of the particular compositions of the present invention employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compositions employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.
- a composition of the disclosure can be administered via one or more routes of administration using one or more of a variety of methods known in the art.
- routes of administration include intravenous, intramuscular, intradermal, intraperitoneal, subcutaneous, spinal or other parenteral routes of administration, for example by injection or infusion.
- parenteral administration means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, epidural and intrasternal injection and infusion.
- a composition can be administered via a non-parenteral route, such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically.
- a non-parenteral route such as a topical, epidermal or mucosal route of administration, for example, intranasally, orally, vaginally, rectally, sublingually or topically.
- the antibody and/or fusion protein can be prepared with carriers that will protect against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems.
- Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen,
- compositions can be administered with medical devices known in the art.
- a therapeutic composition of the disclosure can be administered with a needleless hypodermic injection device, such as the devices disclosed in U.S. Pat. No. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824; or 4,596,556.
- a needleless hypodermic injection device such as the devices disclosed in U.S. Pat. No. 5,399,163; 5,383,851; 5,312,335; 5,064,413; 4,941,880; 4,790,824; or 4,596,556.
- Examples of well-known implants and modules useful in the present disclosure include: U.S. Pat. No. 4,487,603, which discloses an implantable micro-infusion pump for dispensing medication at a controlled rate; U.S. Pat. No. 4,486,194, which discloses a therapeutic device for
- U.S. Pat. No. 4,447,233 which discloses a medication infusion pump for delivering medication at a precise infusion rate
- U.S. Pat. No. 4,447,224 which discloses a variable flow implantable infusion apparatus for continuous drug delivery
- U.S. Pat. No. 4,439,196 which discloses an osmotic drug delivery system having multi-chamber compartments
- U.S. Pat. No. 4,475,196 which discloses an osmotic drug delivery system.
- Many other such implants, delivery systems, and modules are known to those skilled in the art.
- an antibody and/or fusion protein described can be admininstered to a subject using a vector, e.g., a viral vector, e.g., using known methods.
- a viral vector can be used to introduce an antibody and/or fusion protein into a cancer cell (e.g., a tumor cell). Introduction of such antibody and/or fusion protein can increase susceptibility to a subject’s immune system and/or one or more additional therapeutic agents (see, e.g., WO2017/075533).
- a nucleic acid sequence encoding an antibody and/or fusion protein described herein can be cloned into a number of types of vectors.
- a nucleic acid can be cloned into a plasmid, a phagemid, a phage derivative, an animal virus, and a cosmid.
- Other vectors can include expression vectors, replication vectors, probe generation vectors, sequencing vectors, and viral vectors.
- the vector can be a foamy viral (FV) vector, a type of retroviral vector made from spumavirus.
- FV foamy viral
- Viral vector design and technology is well known in the art as described in Sambrook et al, (Molecular Cloning: A Laboratory Manual, 2001), and in other virology and molecular biology manuals.
- viral vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, lentiviruses, poxviruses, herpes simplex 1 virus, herpes virus, oncoviruses (e.g., murine leukemia viruses), and the like.
- a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers, (e.g., WO 01/96584; WO 01/29058; and U.S. Pat. No. 6,326,193).
- Lentiviral and Retroviral transduction can be enhanced by the addition of polybrene (SantaCruz sc-134220; Millipore TR- 1003-G; Sigma 107689), a cationic polymer (also known as hexamehtrine bromide) that is used to increase the efficiency of the retrovirus transduction.
- Retroviruses are enveloped viruses that belong to the viral family Retroviridae.
- the virus replicates by using a viral reverse transcriptase enzyme to transcribe its RNA into DNA.
- the retroviral DNA replicates as part of the host genome, and is referred to as a provirus.
- a selected gene can be inserted into a vector and packaged in retroviral particles using techniques known in the art.
- the recombinant virus can then be isolated and delivered to cells of the subject in vivo.
- retroviral systems are known in the art, (see, e.g., U.S. Pat Nos. 5,994,136, 6,165, 782, and 6,428,953).
- Retroviruses include the genus of Alpharetrovirus (e.g., avian leukosis virus), the genus of Betaretrovirus; (e.g., mouse mammary tumor virus) the genus of Deltaretrovirus (e.g., bovine leukemia virus and human T-lymphotropic virus), the genus of Epsilonretrovirus (e.g., Walleye dermal sarcoma virus), and the genus of Lentivirus.
- a retrovirus is a lentivirus a genus of viruses of the Retroviridae family, e.g., characterized by a long incubation period.
- Lentiviruses are unique among the retroviruses in being able to infect non- dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so can be used as an efficient gene delivery vector.
- a lentivirus can be, but not limited to, human immunodeficiency viruses (HIV-1 and HIV-2), simian
- S1V immunodeficiency virus
- FMV feline immunodeficiency virus
- EIA equine infections anemia
- visna virus vectors derived from lentiviruses offer the means to achieve significant levels of gene transfer in vivo.
- a vector is an adenovirus vector.
- Adenoviruses are a large family of viruses containing double stranded DNA. They replicate the DNA of the host cell, while using a host’s cell machinery to synthesize viral RNA DNA and proteins. Adenoviruses are known in the art to affect both replicating and non-replicating cells, to accommodate large transgenes, and to code for proteins without integrating into the host cell genome.
- an AAVP vector is used.
- An AAVP vector is a hybrid of prokaryotic-eukaryotic vectors, which are chimeras of genetic cis-elements of recombinant adeno-associated virus and phage.
- An AAVP combines selected elements of both phage and AAV vector systems, providing a vector that is simple to produce in bacteria and can exhibit little or no packaging limit, while allowing infection of mammalian cells combined with integration into the host chromosome.
- Vectors containing many of the appropriate elements are commercially available, and can be further modified by standard methodologies to include the necessary sequences.
- AAVPs do not require helper viruses or trans-acting factors.
- a human papilloma (HPV) pseudovirus is used.
- DNA plasmids can be packaged into papillomavirus LI and L2 capsid protein to generate pseudovirion that can efficiently deliver DNA.
- the encapsulation can protect the DNA from nucleases and provides a targeted delivery with a high level of stability.
- Many of the safety concerns associated with the use of viral vectors can be mitigated with an HPV pseudovirus.
- Other methods and examples are in Hung, C., et al.,Plos One, 7:7(e40983); 2012, U.S. Patent
- an oncolytic virus is used.
- Oncolytic virus therapy can selectively replicate the virus in cancer cells, and can subsequently spread within a tumor, e.g., without affecting normal tissue.
- an oncolytic virus can preferentially infect and kill cells without causing damage to normal tissues.
- Oncolytic viruses can also effectively induce immune responses to themselves as well as to the infected tumor cell.
- oncolytic viruses fall into two classes: (I) viruses that naturally replicate preferentially in cancer cells and are nonpathogenic in humans.
- Exemplary class (I) oncolytic viruses include autonomous parvoviruses, myxoma virus (poxvirus), Newcastle disease virus (NDV;
- a second class (II) includes viruses that are genetically manipulated for use as vaccine vectors, including measles virus (paramyxovirus), poliovirus (picornavirus), and vaccinia virus (poxvirus). Additionally, oncolytic viruses may include those genetically engineered with mutations/deletions in genes required for replication in normal but not in cancer cells including adenovirus, herpes simplex virus, and vesicular stomatitis virus. Oncolytic viruses can be used as a viral transduction method due to their low probability of genetic resistance because they can target multiple pathways and replicate in a tumor-selective method. The viral dose within a tumor can increase over time due to in situ viral amplification (as compared to small molecule therapies which decrease with time), and safety features can be built in (i.e., drug and immune sensitivity).
- an antibody and/or fusion protein as described herein may be included in a course of treatment that further includes administration of at least one additional agent to a subject.
- an additional agent administered in combination with an antibody and/or fusion protein as described herein can be cytarabine (cytosine arabinoside, or ara-C) and/or antharcycline drugs such as doxorubicin, daunorubicin, daunomycin, idarubicin and mitoxantrone; other chemotherapeutic drugs such as Hydroxyurea (Hydrea®), Decitabine (Dacogen®), Cladribine (Leustatin®, 2-CdA), Fludarabine (Fludara®), Topotecan, Etoposide (VP- 16), 6-thioguanine (6-TG), corticosteroid drugs, such as prednisone or dexamethasone (Decadron®), methotrexate (MTX), 6-mer
- a fusion protein described herein e.g., a fusion protein consisting of or comprising one or more VHH and one or more tumor antigen
- a cellular therapeutic e.g., a CAR-T cell
- antibody drug conjugate that binds to such one or more tumor antigen as described in, e.g., WO2017/075537,
- an additional agent administered in combination with an antibody and/or fusion protein as described herein may be administered at the same time as an antibody and/or fusion protein, on the same day as an antibody and/or fusion protein, or in the same week as an antibody and/or fusion protein.
- an additional agent administered in combination with an antibody and/or fusion protein as described herein may be administered in a single formulation with an antibody and/or fusion protein.
- an additional agent administered in a manner temporally separated from administration of an antibody and/or fusion protein as described herein e.g., one or more hours before or after, one or more days before or after, one or more weeks before or after, or one or more months before or after administration of an antibody and/or fusion protein.
- the administration frequency of one or more additional agents may be the same as, similar to, or different from the administration frequency of an antibody and/or fusion protein as described herein.
- compositions When compositions are to be used in combination with a second active agent, the compositions can be co-formulated with the second agent or the compositions can be formulated separately from the second agent formulation.
- the respective pharmaceutical compositions can be mixed, e.g., just prior to administration, and administered together or can be administered separately, e.g., at the same or different times.
- combined administration of an antibody and/or fusion protein described herein and an additional agent results in an improvement in a condition or a symptom thereof to an extent that is greater than one produced by either the antibody (or fusion protein) or the additional agent alone.
- the difference between the combined effect and the effect of each agent alone can be a statistically significant difference.
- combined administration of an antibody and/or fusion protein described herein and an additional agent allows administration of the additional agent at a reduced dose, at a reduced number of doses, and/or at a reduced frequency of dosage compared to a dosing regimen for the additional agent, e.g., a standard dosing regimen approved for the additional agent.
- kits can be provided in a kit.
- the kit includes (a) a container that contains an antibody and/or fusion protein described herein (e.g., a pharmaceutical composition comprising an antibody and/or fusion protein described herein) and, optionally (b) informational material.
- the informational material can be descriptive, instructional, marketing or other material that relates to the methods described herein and/or the use of an antibody and/or fusion protein, e.g., for therapeutic benefit.
- the informational material of a kit is not limited in its form.
- the informational material can include information about production of an antibody and/or fusion protein, amino acid of an antibody and/or fusion protein, nucleic acid encoding an antibody and/or fusion protein, molecular weight of an antibody and/or fusion protein, concentration, date of expiration, batch or production site information, and so forth.
- the informational material relates to methods of administering an antibody and/or fusion protein, e.g., in a suitable amount, manner, or mode of administration (e.g., a dose, dosage form, or mode of administration described herein).
- the method can be a method of treating a subject having a CLL-1 associated disorder.
- the informational material e.g., instructions
- the informational material can also be provided in other formats, such as Braille, computer readable material, video recording, or audio recording.
- the informational material of the kit is contact information, e.g., a physical address, email address, website, or telephone number, where a user of the kit can obtain substantive information about an antibody therein and/or their use in the methods described herein.
- the informational material can also be provided in any combination of formats.
- kits can include other ingredients, such as a solvent or buffer, a stabilizer, or a preservative.
- a kit can also include other agents, e.g., a second or third agent, e.g., other therapeutic agents.
- the components can be provided in any form, e.g., liquid, dried or lyophilized form.
- the components can be substantially pure (although they can be combined together or delivered separate from one another) and/or sterile.
- the liquid solution can be an aqueous solution, such as a sterile aqueous solution.
- reconstitution generally is by the addition of a suitable solvent.
- the solvent e.g., sterile water or buffer, can optionally be provided in the kit.
- a kit can include one or more containers for an antibody (and/or fusion protein) and/or other agents.
- a kit contains separate containers, dividers or compartments for an antibody and/or fusion protein and informational material.
- an antibody and/or fusion protein can be contained in a bottle, vial, or syringe, and the informational material can be contained in a plastic sleeve or packet.
- the separate elements of a kit are contained within a single, undivided container.
- an antibody and/or fusion protein can be contained in a bottle, vial or syringe that has attached thereto the informational material in the form of a label.
- a kit can include a plurality (e.g., a pack) of individual containers, each containing one or more unit dosage forms (e.g., a dosage form described herein) of an antibody and/or fusion protein.
- Containers can include a unit dosage, e.g., a unit that includes an antibody and/or fusion protein.
- a kit can include a plurality of syringes, ampules, foil packets, blister packs, or medical devices, e.g., each containing a unit dose.
- the containers of kits can be air tight, waterproof (e.g., impermeable to changes in moisture or evaporation), and/or light-tight.
- a kit can optionally include a device suitable for administration of an antibody and/or fusion protein, e.g., a syringe or other suitable delivery device.
- a device can be provided preloaded with an antibody and/or fusion protein, e.g., in a unit dose, or can be empty, but suitable for loading.
- Recombinant expression of a gene can include construction of an expression vector containing a nucleic acid that encodes the polypeptide.
- a vector for the production of the polypeptide can be produced by recombinant DNA technology using techniques known in the art.
- Known methods can be used to construct expression vectors containing polypeptide coding sequences and appropriate transcriptional and translational control signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination.
- An expression vector can be transferred to a host cell by conventional techniques, and the transfected cells can then be cultured by conventional techniques to produce
- viral vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, lentiviruses, poxviruses, herpes simplex 1 virus, herpes virus, oncoviruses (e.g., murine leukemia viruses, vaccinia virus), and the like.
- a suitable vector contains an origin of replication functional in at least one organism, a promoter sequence, convenient restriction endonuclease sites, and one or more selectable markers, (e.g., WO 01/96584; WO 01/29058; and U.S. Pat. No.
- lymphocytes e.g., T cells
- lymphocytes e.g., T cells
- An oncolytic viral vector e.g., adenovirus, vaccinia, AAV
- a tumor cell e.g., which can secrete an antibody described herein. See, e.g., WO2017/075533.
- CLEC12A also known as CLL-1
- Plates were coated with 1 pg/mL human CLEC12A in PBS (O/N 4C), then blocked with 5% milk/PBST (2hr, room temp); added bacterially expressed sdAbs diluted in 1 : 1 in Block (lhr), washed 5X with PBST, detected with mouse anti- myc-Tag monoclonal antibody (mAb) for lhr, followed by goat anti-mouse IgG-HRP (lhr), both in Block, washed 5X with PBST between antibodies, and plate developed 30 minutes. Each plate included 4 controls / 92 samples per plate. About 32% of sdAb screened were ELISA positive. Figure 1 shows binding data for certain clones.
- U937 cells (2.5c10 L 5) were Fc blocked (human Fc block, BD Cat. # BDB564220) for lOmin at room temperature, then washed once with FACS buffer (PBS + 1% BSA + 0.1% Sodium Azide) by spinning at 500G for 2min; VHH clones (3pg/ml as final cone) in FACS buffer were added to the cell pellets and incubated for 30min at 4°C, then washed twice with FACS buffer by spinning at 500G for 2min; anti-His-PE (R&D Systems, Cat.
- VHH clones (3ug/ml as starting conc./test, with 3x serial dilutions) in FACS buffer were added to Fc blocked U937 or 293T-CLL1 cells (2.5c10 L 5), the mixtures were incubated for 30min at 4°C, then washed twice with FACS buffer by spinning at 500G for 2min; anti-His-PE was added to cells, the mixture was incubated for 30min at 4°C and washed twice with FACS buffer by spinning at 500G for 2min; 1% PFA (Paraformaldehyde) in PBS was added to cell pellet to fix the cells, followed by FACS analysis.
- VHH clones were assessed for binding to E1937. Briefly, VHH clones
- ELISAs were used to determine if the VHH clones recognized distinct or similar epitopes.
- 96 well plates (Pierce Cat. # 15041) were coated with 0. lug/ml soluble CLEC12a-His (Sino Biological # 11896-H07H) in 0.1 M carbonate buffer, pH9.5, O/N at 4C.
- the plate was blocked with 0.3% nonfat milk in TBS (200 ul/well) for 1 hr at RT.
- the plates were washed 3x with lxTBST (0.1M Tris, 0.5M NaCl and 0.05% Tween-20).
- the test anti-CLEC12a VHHs were added at 10 ug/ml horizontally across the plate, 100 ul per well and incubated for lh.
- test anti-CLEC12a biotinylated VHHs were added at 0.2 ug/ml vertically down the plate, 100 ul per well and incubate for 1 h.
- HRP- conjugated streptavidin (Pierce #21130) at 1 :2000 (100 ul/well) was added, followed by incubation at RT in the dark for 1 hr.
- 1-Step Ultra TMB-ELISA reagent (Thermo Fisher #34028) was added at 100 ul per well and read at 450 nm. The mapping results indicated that most anti-CLEC12A VHHs recognized the same epitope, except for 1H1 and perhaps 2C2.
- VHHs from 1B1, IG6 and 2F5 recognize the same epitope as the VHHs from clones that differ in CDR1 (i.e., 1 A10 and 2H3).
- CDR1 i.e. 1 A10 and 2H3
- the IH1 VHH was distinct from these, and 2C2 was intermediate. Results are depicted in Figures 7A-7D.
- Epitopes are either linear or conformational.
- a linear epitope is composed of a linear stretch of amino acids in the sequence that does not form 3 -dimensional structure.
- a conformational epitope is one that requires tertiary folding to create the proper binding region.
- Epitope mapping techniques are used for identifying either linear or conformational epitopes. However, linear techniques do not map conformational epitopes. Linear techniques include peptide array scanning, scanning mutagenesis, peptide library phage display and related techniques that define the amino acids found in the linear epitope.
- These linear techniques are used to map the epitope of antibodies that can bind to a non-conformational part of the antigen, e.g., as is known for antibodies that can bind to denatured proteins that have had their conformations disrupted.
- Most epitopes are conformational and are defined by using one or more techniques including X-ray co-crystallography, conformational peptide scanning as can be performed with large peptides displayed on the surface of phage, mutagenesis technologies whereby specific amino acid residues of the antigen are mutated or changed (often to alanine) and the presence of the complex is detected (often with fluorescence, e.g., in FRET assays), a technique that can be automated, i.e., as when many plasmid clones are generated in a library format using computers to perform statistical calculations of the database, cross-link coupled Mass Spectrometry in which the antibody and the antigen are tagged with a mass-labeled chemical crosslinker.
- the antibody/antigen complex is confirmed using high mass MALDI detection. Once created the antibody/antigen complex is extremely stable, and various enzymes and digestion conditions are applied to the complex to provide many different overlapping peptides. Identification of these peptides is performed using high-resolution mass spectrometry and MS/MS techniques. Identification of the crosslinked peptides is determined using mass tags linked to the cross-linking reagents. After MS/MS fragmentation and data analysis using specific interaction software, both epitope and paratope are determined in the same experiment. In another technique, Hydrogen Deuterium Exchange (HDX), the availability of hydrogen molecules in the backbone of a protein structure is measured.
- HDX Hydrogen Deuterium Exchange
- both the unbound antigen and the bound antibody-antigen complex are incubated in deuterated water in order to exchange any hydrogens from exposed amino acids of the protein's backbone.
- residues of the epitope are determined.
- One or more of these technologies is used to identify linear and conformational epitopes of the VHH antibodies of the invention.
- 11896-H07H was run on a 4-12% gradient SDS gel (Invitrogen, NP0321) under reducing conditions. After blotting, using the iBlot 2 system (Invitrogen, IB23002), the membrane was blocked in 5% non-fat milk in Tris buffered saline (TBS) for 1 hr. Next, half the blot was incubated with about 1 ug/ml protein supernatant of myc tagged anti CLEC12a VHH for 2 hrs followed by 3x wash in wash buffer (lx TBST: 0.1 M Tris, 0.5 M NaCl, 0.05% Tween 20).
- TBS Tris buffered saline
- the blot was then incubated with 1 :2000 HRP-anti-Myc (Rockland/Fisher, 50-105-8097) for another 1 hr and washed 3x in wash buffer and then developed using SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Fisher, 34095). The other half of the blot was directly incubated with 1 :2000 HRP-anti-his (BioLegend/Fisher, 652504) for 1 hr at 0.1 pg/ml and developed as a positive control for the presence of Clecl2A on the blot. Only one clone, VHH clone 1H1, was found to bind denatured Clecl2A by Western Blot. Results are depicted in Figure 8.
- VHH clones were tested for their ability to compete for binding with a known anti-Clecl2A scFv.
- a fusion protein comprising the anti-Clecl2A scFv SC02-357 described in US Patent 7741443 (included herein as SEQ ID NO:26) was immobilized on plastic plates followed by binding of recombinant Clecl2A.
- Clecl2a proteins was tested by ELISA. Briefly, a 96 well plate (Pierce, 15041) was coated with l.O ug/ml Clecl2A (Sino Biological, 11896-H07H or ABclonal, RP01018) in 0.1 M carbonate, pH9.5 overnight at 4°C. The plate was blocked with 200 m ⁇ /well 0.3% non-fat milk in Tris buffered saline (TBS) for 1 hr at room temperature (RT). Then, the plates were washed 3x with wash buffer (lx TBST: 0.1 M Tris, 0.5 M NaCl, 0.05% Tween 20).
- TBS Tris buffered saline
- the plate was coated with the Clecl2A proteins as above, but instead of probing with the 2H3 VHH, a 2H3 VHH-G4Sx4-CD19-His fusion protein was added starting at 5 pg/ml with 3 fold dilutions. Also, instead of the anti-Myc reagent, the anti-CD 19 Ab FMC63 was added at 1 pg/ml cone. After washing, the protocol using the HRP anti-mlgG reagent was followed as above. Graphs were generated using the Softmax software and are shown in Figure 10B.
- Clecl2A is a Type II membrane protein, i.e. the C-terminus is extracellular.
- Figure 4A the Clecl2A’s from each source were also immobilized using an anti-Clecl2A antibody immobilized on plastic plates first, which gave similar results ( Figure 10B).
- Figure 10B the amino acid sequences of the two recombinant Clecl2A proteins.
- the canonical sequence (UniProt; Q5QGZ9-1) contains a Lysine (K in bold underline; SEQ ID NO:27 ).
- NP OOl 193939.1, isoform3 or a cDNA from Genscript both of which contain a Lysine residue at amino acid 254.
- K/Q amino-acid is within or close to the 2H3 epitope, and is consistent with the 2H3 epitope being close to the C-terminus.
- scFv binds to both protein variants.
- Figure 11 A shows the amino acids crosslinked between 2H3 and Clecl2A defining the region of the 2H3 epitope.
- Amino acids 251 (T); 252 (Y); and 260 (K) (based on full length Clecl2A SEQ ID NO 28) were identified as cross linked.
- the epitope spans the region containing the K/Q variant, entirely consistent with a change in charge in this region (K vs. Q) disrupting the 2H3 epitope as suggested in Example 3.
- Figure 1 IB shows the epitope for the SC02-357 scFv. Again, its presence near the C-terminal is consistent with all other results, i.e., a distinct epitope from 2H3, yet within the last 50 AA of Clecl2A.
- CleclA and Clecl2B were the most related with identity in their extracellular domains of 36% and 32%, respectively.
- Fc-blocked 293T-Clecl2A; Clecl2B; or CleclA cells were resuspended in 50 m ⁇ FACs buffer (PBS / 1% BSA / 0.1% Sodium Azide) and then added to 50 m ⁇ of a 2H3VHH-CD19-His fusion protein (His Trap Excel (GE Healthcare, 17-3712 purified) serially diluted in FACS buffer, starting at 5 pg/ml, final concentration.
- the sequence of the 2H3 VHH used is a version of SEQ ID NO: 15 lacking the C-terminal myc and His tags disclosed in SEQ ID 29Serial dilutions were done in 3-fold steps.
- the cells and the 2H3VHH-CD19 fusion protein were incubated together for 30 minutes at 4°C.
- the samples were washed in FACS buffer and centrifuged at 500 RCF at 4°C for 2 minutes. This wash step was repeated and then the cells were resuspended in FACs buffer containing 2.5 pg/ml PE-labeled FMC63 (MilliporeSigma, MAB1794H). After incubation for 30 minutes at 4°C, the cells were washed twice as described resuspended in 150 m ⁇ PBS / 1% paraformaldehyde to fix the cells.
- Leu Glu Phe lie Lys Ser Gin Ser Arg Ser Tyr Asp Tyr Trp Leu Gly
- SEQ ID NO:3 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids ( TSGPGGQGA ), (ii) a myc-tag
- SEQ ID NO: 4 (underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- SEQ ID NO: 5 underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- SEQ ID NO: 6 underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 8 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids ( TSGPGGQGA ), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 9 (underlining denotes CDRL CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 10 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 12 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids ( TSGPGGQGA ), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 13 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 14 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 15 (underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag (EQKLISEEDL), (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 16 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids ( TSGPGGQGA ), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 17 (underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 18 (underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO: 19 (underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) : OVOLOESGGGLVOAGGSLRLSCVVSGATSNVNAMGWYROAPGKERELVAAISSGGSTS YRDSVKGRFTISRDNAKNTLYLOMNSLKPEDTAMYYCAAODWATEGYEYDYWGOGT L VT VS S TSGPGGQGAEQKLISEEDLGAHHHHHHGAS
- SEQ ID NO:20 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids ( TSGPGGQGA ), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO:21 underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO:22 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO:23 underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) : OVOLOEFGGGLVOLGGSARLSCVVSGNMLDLNTMAWYROGELVAALGISTYYAESVK GRFTISRDNAKNTLYLQMN SLKSEDT AVYY C ARDYNFESW GQGTL VT V S S TSGPGGQG AEQKLISEEDLGAHHHHGAS
- SEQ ID NO:24 (underlining denotes CDR 1. CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids ( TSGPGGQGA ), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
- SEQ ID NO:25 underlining denotes CDRE CDR2. CDR3. sequentially; bolded italics at C- ter minus denotes (i) a linker of 9 amino acids (TSGPGGQGA), (ii) a myc-tag
- EQKLISEEDL (iii) a linker of 2 amino acids (GA), (iv) a hexa-histidine tag (HHHHHH), and (v) an additional 3 amino acids (GAS)) :
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- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Life Sciences & Earth Sciences (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
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Abstract
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Priority Applications (9)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020217020449A KR20210124201A (en) | 2018-11-30 | 2019-11-27 | Single domain antibody to CLL-1 |
EP19888773.9A EP3886903A4 (en) | 2018-11-30 | 2019-11-27 | Single domain antibodies against cll-1 |
CA3121478A CA3121478A1 (en) | 2018-11-30 | 2019-11-27 | Single domain antibodies against cll-1 |
MX2021006362A MX2021006362A (en) | 2018-11-30 | 2019-11-27 | Single domain antibodies against cll-1. |
JP2021530988A JP2022511799A (en) | 2018-11-30 | 2019-11-27 | Single domain antibody against CLL-1 |
AU2019390483A AU2019390483A1 (en) | 2018-11-30 | 2019-11-27 | Single domain antibodies against CLL-1 |
US17/298,246 US20220112292A1 (en) | 2018-11-30 | 2019-11-27 | Single domain antibodies against cll-1 |
CN201980089865.0A CN113507937A (en) | 2018-11-30 | 2019-11-27 | Single domain antibodies to CLL-1 |
IL283476A IL283476A (en) | 2018-11-30 | 2021-05-26 | Single domain antibodies against cll-1 |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
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US201862774025P | 2018-11-30 | 2018-11-30 | |
US62/774,025 | 2018-11-30 | ||
US201862780092P | 2018-12-14 | 2018-12-14 | |
US62/780,092 | 2018-12-14 |
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WO2020113063A1 true WO2020113063A1 (en) | 2020-06-04 |
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Application Number | Title | Priority Date | Filing Date |
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PCT/US2019/063691 WO2020113063A1 (en) | 2018-11-30 | 2019-11-27 | Single domain antibodies against cll-1 |
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US (1) | US20220112292A1 (en) |
EP (1) | EP3886903A4 (en) |
JP (1) | JP2022511799A (en) |
KR (1) | KR20210124201A (en) |
CN (1) | CN113507937A (en) |
AU (1) | AU2019390483A1 (en) |
CA (1) | CA3121478A1 (en) |
IL (1) | IL283476A (en) |
MA (1) | MA54312A (en) |
MX (1) | MX2021006362A (en) |
WO (1) | WO2020113063A1 (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020227073A1 (en) * | 2019-05-04 | 2020-11-12 | Inhibrx, Inc. | Clec12a-binding polypeptides and uses thereof |
CN115772222A (en) * | 2022-12-16 | 2023-03-10 | 浙江康佰裕生物科技有限公司 | anti-CLL 1 single domain antibody and application thereof |
WO2023201238A1 (en) * | 2022-04-11 | 2023-10-19 | Vor Biopharma Inc. | Binding agents and methods of use thereof |
Citations (5)
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US20100285037A1 (en) * | 2007-10-17 | 2010-11-11 | Arie Abo | Antibodies to cll-1 |
WO2015150815A1 (en) * | 2014-04-03 | 2015-10-08 | The University Court Of The University Of Aberdeen | Methods and materials relating to auto-immune disease |
US20160368994A1 (en) * | 2015-06-16 | 2016-12-22 | Genentech, Inc. | Anti-cll-1 antibodies and methods of use |
WO2017091615A1 (en) * | 2015-11-24 | 2017-06-01 | Cellerant Therapeutics, Inc. | Humanized anti-cll-1 antibodies |
US20180273633A1 (en) * | 2012-05-07 | 2018-09-27 | Cellerant Therapeutics, Inc. | Antibodies specific for cll-1 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
MX2017003022A (en) * | 2014-09-12 | 2017-05-12 | Genentech Inc | Anti-cll-1 antibodies and immunoconjugates. |
CN108866088B (en) * | 2018-06-20 | 2023-06-09 | 上海恒润达生生物科技股份有限公司 | Targeting CLL-1 chimeric antigen receptor and uses thereof |
-
2019
- 2019-11-27 EP EP19888773.9A patent/EP3886903A4/en active Pending
- 2019-11-27 MA MA054312A patent/MA54312A/en unknown
- 2019-11-27 US US17/298,246 patent/US20220112292A1/en active Pending
- 2019-11-27 CA CA3121478A patent/CA3121478A1/en active Pending
- 2019-11-27 CN CN201980089865.0A patent/CN113507937A/en active Pending
- 2019-11-27 MX MX2021006362A patent/MX2021006362A/en unknown
- 2019-11-27 JP JP2021530988A patent/JP2022511799A/en active Pending
- 2019-11-27 WO PCT/US2019/063691 patent/WO2020113063A1/en unknown
- 2019-11-27 KR KR1020217020449A patent/KR20210124201A/en unknown
- 2019-11-27 AU AU2019390483A patent/AU2019390483A1/en active Pending
-
2021
- 2021-05-26 IL IL283476A patent/IL283476A/en unknown
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100285037A1 (en) * | 2007-10-17 | 2010-11-11 | Arie Abo | Antibodies to cll-1 |
US20180273633A1 (en) * | 2012-05-07 | 2018-09-27 | Cellerant Therapeutics, Inc. | Antibodies specific for cll-1 |
WO2015150815A1 (en) * | 2014-04-03 | 2015-10-08 | The University Court Of The University Of Aberdeen | Methods and materials relating to auto-immune disease |
US20160368994A1 (en) * | 2015-06-16 | 2016-12-22 | Genentech, Inc. | Anti-cll-1 antibodies and methods of use |
WO2017091615A1 (en) * | 2015-11-24 | 2017-06-01 | Cellerant Therapeutics, Inc. | Humanized anti-cll-1 antibodies |
Non-Patent Citations (1)
Title |
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See also references of EP3886903A4 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2020227073A1 (en) * | 2019-05-04 | 2020-11-12 | Inhibrx, Inc. | Clec12a-binding polypeptides and uses thereof |
WO2023201238A1 (en) * | 2022-04-11 | 2023-10-19 | Vor Biopharma Inc. | Binding agents and methods of use thereof |
CN115772222A (en) * | 2022-12-16 | 2023-03-10 | 浙江康佰裕生物科技有限公司 | anti-CLL 1 single domain antibody and application thereof |
CN115772222B (en) * | 2022-12-16 | 2024-03-29 | 浙江康佰裕生物科技有限公司 | anti-CLL 1 single domain antibodies and uses thereof |
Also Published As
Publication number | Publication date |
---|---|
EP3886903A4 (en) | 2022-07-13 |
CA3121478A1 (en) | 2020-06-04 |
IL283476A (en) | 2021-07-29 |
KR20210124201A (en) | 2021-10-14 |
CN113507937A (en) | 2021-10-15 |
AU2019390483A1 (en) | 2021-07-15 |
US20220112292A1 (en) | 2022-04-14 |
MA54312A (en) | 2022-03-09 |
JP2022511799A (en) | 2022-02-01 |
MX2021006362A (en) | 2021-10-13 |
EP3886903A1 (en) | 2021-10-06 |
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