Conjugates enhancing total cellular accumulation

Abstract

The present description relates to a conjugated compound an antibody covalently linked to enhancer moiety composed by a nuclear localization sequence (NLS), covalently linked to a sterol variant, such as cholic acid (ChAc) or a variant thereof. The enhancer moiety as encompassed herein is able to induce endosome escape of the compound-conjugates by direct membrane destabilization or indirectly by ROS and ceramide production which destabilize endosome-lysosome membrane. The conjugated compound can further comprise payload.

1 .- 20 . (canceled) 
     
     
         21 . A conjugated compound comprising an antibody or an epitope-binding fragment thereof, covalently linked to a peptide comprising a nuclear localization sequence (NLS) other than the SV40 NLS set forth in SEQ ID NO: 1, said peptide covalently linked to a bile acid. 
     
     
         22 . The conjugated compound of  claim 21 , wherein the NLS comprises a monopartite NLS. 
     
     
         23 . The conjugated compound of  claim 21 , wherein the NLS comprises a bipartite NLS. 
     
     
         24 . The conjugated compound of  claim 21 , wherein the NLS comprises a non-classical NLS, wherein the non-classical NLS comprises a hydrophobic and basic PY-NLS. 
     
     
         25 . The conjugated compound of  claim 21 , wherein the nuclear localization signal comprises a/an: PQBP1 NLS (SEQ ID NO: 2), hnRNPA1 NLS (SEQ ID NO: 3), GWG-SV40 NLS (SEQ ID NO: 4), NLS2 RPS1 (SEQ ID NO: 5), NLS1 RPS17 (SEQ ID NO: 6), NLS3 RPS17 (SEQ ID NO: 7), nucleoplasmin NLS (SEQ ID NO: 8), cMyc NLS (SEQ ID NO: 9), TUS NLS (SEQ ID NO: 10), hnRNP D NLS (SEQ ID NO: 11), hnRNP M NLS (SEQ ID NO: 12), HuR NLS (SEQ ID NO: 13), or NLS2-RG RPS17 (SEQ ID NO: 14). 
     
     
         26 . The conjugated compound of  claim 25 , wherein the NLS comprises a variant of the NLS of any one of SEQ ID NOs: 2-14, wherein the variant has nuclear localization activity. 
     
     
         27 . The conjugated compound of  claim 21 , wherein the bile acid comprises: cholic acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid (CDCA), lithocholic acid (LCA), ursodeoxycholic acid (UDCA), glycocholic acid (GCA), glycochenodeoxycholic acid (GCDCA), glycodeoxycholic acid (GDCA), or glycoursodeoxycholic acid (GUDCA). 
     
     
         28 . The conjugated compound of  claim 21 , wherein the bile acid comprises a variant of CA, DCA, CDCA, LCA, UDCA, GCA, GCDCA, GDCA, or GUDCA, wherein the variant triggers ceramide accumulation on the inner leaflet of endosomes and/or triggers increased acid sphingomyelinase (ASM)-mediated cleavage of sphingomyelin to form ceramide. 
     
     
         29 . The conjugated compound of  claim 21 , wherein the bile acid comprises a primary bile acid. 
     
     
         30 . The conjugated compound of  claim 21 , wherein the bile acid comprises a secondary bile acid. 
     
     
         31 . The conjugated compound of  claim 21 , wherein epitope-binding fragment comprises Fv, F(ab′), or F(ab′) 2 . 
     
     
         32 . The conjugated compound of  claim 21 , further comprising a payload covalently linked to the antibody or epitope-binding fragment thereof. 
     
     
         33 . The conjugated compound of  claim 32 , wherein the payload comprises or consists of a radionuclide. 
     
     
         34 . The conjugated compound of  claim 33 , wherein the radionuclide is at least one of  47 Sc,  51 Cr,  52 mMn,  55 Co,  58 Co,  52 Fe,  56 Ni,  57 Ni,  61 Cu,  62 Cu,  64 Cu,  67 Cu,  66 Ga  68 Ga,  67 Ga,  72 As,  77 As,  89 Zr,  90 Y,  94m Tc,  99m Tc,  97 Ru,  105 Rh,  109 Pd,  111 Ag,  110 In,  111 In,  113m In,  114m In,  117m Sn,  121 Sn,  127 Te,  142 Pr,  143 Pr,  149 Pm,  151 Pm,  149 Tb,  153 Sm,  157 Gd,  161 Tb,  166 Ho,  165 Dy,  169 Er,  169 Yb,  175 Yb,  172 Tm,  177 Lu,  186 Re,  188 Re,  191 Pt,  197 Hg,  198 Au,  199 Au,  201 Tl,  203 Pb,  211 At,  212 Bi,  213 Bi,  11 C,  75 Br,  76 Br,  77 Br,  82 Br,  18 F,  120 I,  123 I,  124 I,  125 I,  131 I,  89 Sr, or  225 Ac. 
     
     
         35 . The conjugated compound of  claim 32 , wherein the payload comprises a small molecule toxin. 
     
     
         36 . The conjugated compound of  claim 32 , wherein the payload comprises a chemotherapeutic agent. 
     
     
         37 . The conjugated compound of  claim 36 , wherein the chemotherapeutic agent comprises a microtubule disrupting agent, a DNA targeting agent, an RNA polymerase inhibitor, a topoisomerase inhibitor, or a DNA alkylated agent. 
     
     
         38 . The conjugated compound of  claim 32 , wherein the payload comprises an imaging molecule. 
     
     
         39 . The conjugated compound of  claim 38 , wherein the imaging molecule comprises a fluorescence molecule or a radionuclide. 
     
     
         40 . A method for treating cancer in a subject, the method comprising:
 (a) providing the conjugated compound of  claim 32 , wherein the antibody or epitope-binding fragment thereof specifically binds to an epitope expressed on the subject's cancer cells, and wherein the payload comprises a chemotherapeutic agent; and   (b) administering the conjugated compound to the subject.