METHODS AND COMPOSITIONS FOR TREATING BETA-THALASSEMIA AND SICKLE CELL DISEASE

Abstract

Compounds, pharmaceutical compositions, and methods for treating anemia -thalassemia anemia or sickle cell anemia.

Claims

1.-27. (canceled)

28. A compound of the formula selected from the group consisting of: Formula (VI), Formula (VII), and Formula (VIII): ##STR00106## or a pharmaceutically acceptable salt thereof, wherein: R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12 and R.sup.13 are each independently selected from the group consisting of hydrogen, optionally substituted alkyl, N(R.sup.B).sub.2, and N(R.sup.A)C(O)R.sup.A; provided that in Formula (VI), R.sup.10 is optionally substituted alkyl, N(R.sup.B).sub.2, or N(R.sup.A)C(O)R.sup.A. R.sup.14 is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, or optionally substituted alkynyl; each instance of R.sup.A is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl; and each instance of R.sup.B is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl;

29. The compound of claim 28, wherein the compound is selected from the group consisting of Formula (VI-a), (VII-a), and (VIII-a): ##STR00107## in which each of R.sup.10, R.sup.11, and R.sup.14 is as defined in claim 28.

30. The compound of claim 29, wherein R.sup.14 is optionally substituted alkyl.

31. The compound of claim 30, wherein R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein: n is 1, 2, 3, 4, or 5; and R.sup.B is independently optionally substituted C.sub.1-6 alkyl.

32. The compound of claim 31, wherein R.sup.14 is (CH.sub.2).sub.2N(CH.sub.3).sub.2.

33. The compound of claim 29, wherein R.sup.10 is N(R.sup.B).sub.2 or N(R.sup.A)C(O)R.sup.A, wherein: R.sup.A is independently hydrogen or optionally substituted alkyl; and R.sup.B is independently hydrogen or optionally substituted alkyl.

34. The compound of claim 33, wherein (i) each R.sup.B is methyl or hydrogen, or (ii) one instance of R.sup.B is optionally substituted C.sub.1-C.sub.5 alkyl and the other instance of R.sup.B is hydrogen.

35. The compound of claim 33, wherein one instance of R.sup.A is optionally substituted alkyl and the other instance of R.sup.A is hydrogen.

36. The compound of claim 29, wherein R.sup.11 is hydrogen.

37. The compound of claim 29, wherein the compound is selected from the group consisting of the compounds of the formula: ##STR00108## or a pharmaceutically acceptable salt thereof.

38. A pharmaceutical composition comprising a compound of claim 28, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

39. A method of inducing globin production, comprising: contacting a cell with an effective amount of a compound of claim 28.

40. The method of claim 39, wherein the contacting step is performed by administering an effective amount of the compound of Formula (VI), Formula (VII), or Formula (VIII), or a pharmaceutically acceptable salt thereof to a subject in need thereof.

41. The method of claim 40, wherein the subject is a human patient who suffers from or is suspected of having anemia.

42. The method of claim 41, wherein the anemia is -thalassemia or sickle cell anemia.

43. The method of claim 40, wherein the compound is administered orally.

44. The method of claim 40, wherein the pharmaceutical composition is administered in combination with an additional therapeutic agent.

45. A method of treating anemia, comprising: administering to a subject in need thereof an effective amount of a compound of claim 28.

46. The method of claim 45, wherein the subject is a human patient who suffers from or is suspected of having -thalassemia or sickle cell anemia.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0104] FIG. 1 shows the structures of S51021, naphthalimides, and SS-2394.

[0105] FIG. 2 shows the structures of five subgroups of N-substituted 1,8-naphthalimide derivatives. The naphthalimide derivatives were prepared by the condensation of naphthalic anhydride derivatives with substituted aniline (A1-A14), dimethylaminopropylamine/dimethylethylene diamine (B1-B4), alkly amine (C1-C5), or ethyl 4-aminobutyrate (D1). The amino-substituted naphthalimide derivatives (E1-E3) were prepared by the reduction of the nitro-substituted naphthalimide compounds A2, B2, and B3 by catalytic hydrogenation.

[0106] FIGS. 3A-3F are a series of graphs showing the dosage effects of six naphthalimide derivatives, Compounds A2, Compound B1, Compound B4, Compound E1, Compound E2, and Compound E3, respectively, in mediating the globin gene induction and cell proliferation in primary human erythroid cells. Six naphthalimide derivatives with identical pharmacophore were selected to estimate their globin gene inducing abilities in primary human erythroid cells. After 3 days of compounds treatment, the total mRNA was extracted and the relative induction fold of the globin gene was analyzed by quantitative RT-PCR. The cell proliferation rate was examined by AlamarBlue reagent. Data are presented as the relative expression level in meanSEM, N=8.

[0107] FIGS. 4A-4D are graphs showing the expression profiles of the globin gene and HbF-related regulators after compounds treatment in the primary human erthroid cells. The primary human erythroid cells were treated with HU (145.9 M), NaB (221.6 M), S51021 (21.8 M) or SS-2394 (2.3 M) at their IC.sub.50 for 3 days. FIG. 4A shows that the total RNA was extracted, and the expression profiles of globin, NF-E4, BCL11A, and c-Myb were analyzed by relative quantitative RT-PCR. Data are presented as the relative expression level in meanSEM, N=8. FIG. 4B shows a graph and two panels. Upper panel: For detection of the induction level of globin, fifty microgram of total cell lysate was analyzed by western blot analysis using antibody against globin, or -actin. Lower panel: The signal intensity was quantified by an image-analyzing software. The intensity of globin expression levels of mock control was set as 1. Data are shown as the meanSEM, N=5. FIG. 4C shows a graph and two panels. Upper panel: For detection of the expression level of BCL11A, thirty microgram of total protein was separated by SDS-PAGE electrophoresis, transferred to PVDF membrane, and immunobloted with antibody against BCL11A and -actin. Lower panel: The signal was quantified by an image-analyzing software. The intensity of the mock control BCL11A levels were set as 1. Data are shown as the meanSEM, N=5. FIG. 4D is a graph showing that the HU-nonresponding primary human erythroid cells were treated with HU (145.9 M), NaB (221.6 M), S51021 (21.8 M) or SS-2394 (2.3 M) for 3 days. Total RNA was extracted, and the expression level of globin gene was analyzed by relative quantitative RT-PCR. Data are shown as the meanSEM, N=3.

[0108] FIG. 5 is a graph showing the phosphorylation status of p38 in primary erythroid cells treating with HU, NaB, S51021, or SS-2394. Upper panel: For detection of the phosphorylation status of p38, twenty microgram of total cell lysate extracted from the primary human erythroid cells treated with HU (145.9 M), NaB (221.6 M), S51021 (21.8 M) or SS-2394 (2.3 M) was analyzed by Western blot analysis using antibody against p-p38 or p38. Lower panel: The signal intensity was quantified by an image-analyzing software. The intensity of p-p38 levels of mock control cells was set as 1. Data are shown as the meanSEM, N=5.

[0109] FIG. 6 is a graph showing the acetylation status of histone H4 in primary erythroid cells treating with HU, NaB, S51021 or SS-2394. Upper pnel: To examine the acetylation pattern of histone H4, the primary human erythroid cells were treated with HU (145.9 M), NaB (221.6 M), S51021 (21.8 M) or SS-2394 (2.3 M) at their IC.sub.50 for 3 days. Total histones were extracted by histone extraction buffer. Thirty microgram of histone extract was used for western blot analysis and immunobloted with antibody against histone H4 or acetylated histone H4. Lower panel: The signal intensity was quantified by an image-analyzing software. The intensity of mock control was set as 1. Data are shown as the meanSEM, N=4.

[0110] FIG. 7 shows two exemplary synthetic schemes, Scheme 1 and Scheme 2, for synthesing the compounds described herein.

[0111] FIG. 8 shows the effect of globin-induction and cytotoxicity against primary erythroid cells by the representative analogues of 2,3-dihydro-1H-benzo[de]isoquinoline.

DETAILED DESCRIPTION OF THE INVENTION

[0112] The present disclosure stems, in part, from the unexpected discovery that a number of small molecules were found to induce globin gene expression. The compounds identified herein are useful for treatment of -thalassemia or sickle cell disease through induction of endogenous embryonic/fetal globin chains. Accordingly, disclosed herein are compounds capable of inducing globin gene expression and uses thereof in treating anemia such as -thalassemia or sickle cell disease.

Compounds Capable of Inducing Globin Gene Expression

[0113] In one embodiment, the compound is represented by structural formula A, I, I-a, II, III, IV, III-a, III-b, (V-a), (VI), (VI-a), (VII), (VII-a), (VIII), (VIII-a), (IX), (IX-a), (X), (X-a), (XI), and/or (XI-a) wherein the values for the variables are as defined herein.

[0114] Compounds of interest include naphthalimides, e.g., compounds that include a naphthalimide scaffold substituted with one or more substituents, which are non-hydrogen. The naphthalimide scaffold of Formula (I) may be substituted at any position 1, 2, 3, 4, 5, 6, and/or 7 shown below.

##STR00007##

[0115] In certain embodiments, positions 3 and 4 of the naphthalimide scaffold are substituted. In certain embodiments, position 3 of the naphthalimide scaffold is substituted. In certain embodiments, positions 3, 4, and 7 of the naphthalimide scaffold are substituted. In certain embodiments, positions 3 and 7 of the naphthalimide scaffold are substituted.

[0116] In certain embodiments, a substituent may contribute to optical isomerism and/or stereo isomerism of a compound. The compound(s) of the present disclosure provided herein include the neutral form, salts, solvates, hydrates, and prodrug forms of a compound. All such forms are embraced by the present disclosure. Thus the compounds described herein include salts, solvates, hydrates, prodrug, and isomer forms thereof, including the pharmaceutically acceptable salts, solvates, hydrates, prodrugs, and isomers thereof. In certain embodiments, a compound may be a metabolized into a pharmaceutically active derivative.

[0117] In certain embodiments, the present disclosure employs a compound of Formula (I-a):

##STR00008##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof,
wherein:

[0118] R.sup.1, R.sup.2, R.sup.3, R.sup.4, R.sup.6a, and R.sup.6b are each independently selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B;

[0119] R.sup.5 is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, or optionally substituted aryl;

[0120] each instance of R.sup.A is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl;

[0121] each instance of R.sup.B is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl, or two R.sup.B are taken together with the intervening atoms to form a heterocycle.

[0122] In some embodiments of Formula I-a, R.sup.6a and R.sup.6b are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (II):

##STR00009##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.1, R.sup.2, R.sup.3, R.sup.4 and R.sup.5 are as described herein.

[0123] As described generally above, R.sup.1 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.1 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.1 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, heteroaryl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.1 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, heteroaryl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.1 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.1 is hydrogen. In some embodiments, R.sup.1 is C.sub.1-6 alkyl. In certain embodiments, R.sup.1 is C(O)OR.sup.A. In certain embodiments, R.sup.1 is CO.sub.2H. In certain embodiments, R.sup.1 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.1 is halogen. In certain embodiments, R.sup.1 is fluoro. In certain embodiments, R.sup.1 is chloro. In certain embodiments, R.sup.1 is bromo. In certain embodiments, R.sup.1 is iodo. In some embodiments, R.sup.1 is N(R.sup.B).sub.2. In some embodiments, R.sup.1 is NH.sub.2. In some embodiments, R.sup.1 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.1 is NHCH.sub.3. In certain embodiments, R.sup.1 is NO.sub.2. In some embodiments, R.sup.1 is optionally substituted naphthyl.

[0124] As described generally above, R.sup.2 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.2 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.2 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.2 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.2 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.2 is hydrogen. In some embodiments, R.sup.2 is C.sub.1-6 alkyl. In certain embodiments, R.sup.2 is C(O)OR.sup.A. In certain embodiments, R.sup.2 is CO.sub.2H. In certain embodiments, R.sup.2 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.2 is halogen. In certain embodiments, R.sup.2 is fluoro. In certain embodiments, R.sup.2 is chloro. In certain embodiments, R.sup.2 is bromo. In certain embodiments, R.sup.2 is iodo. In some embodiments, R.sup.2 is NH.sub.2. In some embodiments, R.sup.2 is N(R.sup.B).sub.2. In some embodiments, R.sup.2 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.2 is NHCH.sub.3. In certain embodiments, R.sup.2 is NO.sub.2. In some embodiments, R.sup.2 is optionally substituted naphthyl.

[0125] As described generally above, R.sup.3 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.3 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.3 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.3 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.3 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.3 is hydrogen. In some embodiments, R.sup.3 is C.sub.1-6 alkyl. In certain embodiments, R.sup.3 is C(O)OR.sup.A. In certain embodiments, R.sup.3 is CO.sub.2H. In certain embodiments, R.sup.3 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.3 is halogen. In certain embodiments, R.sup.3 is fluoro. In certain embodiments, R.sup.3 is chloro. In certain embodiments, R.sup.3 is bromo. In certain embodiments, R.sup.3 is iodo. In some embodiments, R.sup.3 is N(R.sup.B).sub.2. In some embodiments, R.sup.3 is NH.sub.2. In some embodiments, R.sup.3 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.3 is NHCH.sub.3. In certain embodiments, R.sup.3 is NO.sub.2. In some embodiments, R.sup.3 is optionally substituted naphthyl.

[0126] As described generally above, R.sup.4 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.4 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.4 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.4 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.4 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.4 is hydrogen. In some embodiments, R.sup.4 is C.sub.1-6 alkyl. In certain embodiments, R.sup.4 is C(O)OR.sup.A. In certain embodiments, R.sup.4 is CO.sub.2H. In certain embodiments, R.sup.4 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.4 is halogen. In certain embodiments, R.sup.4 is fluoro. In certain embodiments, R.sup.4 is chloro. In certain embodiments, R.sup.4 is bromo. In certain embodiments, R.sup.4 is iodo. In some embodiments, R.sup.4 is N(R.sup.B).sub.2. In some embodiments, R.sup.4 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.4 is NHCH.sub.3. In certain embodiments, R.sup.4 is NO.sub.2. In some embodiments, R.sup.4 is optionally substituted naphthyl.

[0127] As described generally above, R.sup.5 is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, or optionally substituted aryl. In certain embodiments, R.sup.5 is hydrogen. In some embodiments, R.sup.5 is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl or optionally substituted aryl. In certain embodiments, R.sup.5 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. In certain embodiments, R.sup.5 is isopropyl, isobutyl, or isoamyl. In certain embodiments, R.sup.5 is isobutyl. In certain embodiments, R.sup.5 is tert-butyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 1, 2, 3, 4, 5, or 6. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, n is 4. In certain embodiments, n is 5. In certain embodiments, n is 6. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nNHR.sup.B. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is unsubstituted C.sub.1-6 alkyl. In certain embodiment, R.sup.5 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(CH.sub.3)R.sup.B, wherein each R.sup.B is independently optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(CH.sub.3)R.sup.B, wherein each R.sup.B is independently unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(CH.sub.3).sub.2. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(CH.sub.2CH.sub.3)R.sup.B, wherein each R.sup.B is independently optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(CH.sub.2CH.sub.3)R.sup.B, wherein each R.sup.B is independently unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is independently selected from the group consisting of methyl, ethyl, isopropyl, isobutyl, isoamyl, and benzyl. In some embodiments, R.sup.5 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is the same. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 1, 2, 3, 4, 5, or 6, and wherein R.sup.B is independently optionally substituted C.sub.1-6 alkyl or two R.sup.B are taken together with the intervening atoms to form a heterocycle with at least 1-4 heteroatoms. In certain embodiments, the heterocycle is a 3-8 membered ring. In certain embodiments, the heterocycle formed is In certain embodiments, R.sup.5 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 1, 2, 3, 4, or 5, and wherein R.sup.B is independently optionally substituted C.sub.1-6 alkyl or two R.sup.B are taken together with the intervening atoms to form a heterocycle. In some embodiments, R.sup.5 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is different. In certain embodiments, R.sup.5 is (CH.sub.2).sub.nNH.sub.2. In certain embodiments, R.sup.5 is (CH.sub.2).sub.sCO.sub.2R.sup.A, wherein s is 1, 2, 3, 4, 5, or 6. In certain embodiments, R.sup.5 is (CH.sub.2).sub.sCO.sub.2H. In some embodiments, s is 1. In some embodiments, s is 2. In some embodiments, s is 3. In some embodiments, s is 4. In some embodiments, s is 5. In some embodiments, s is 6. In certain embodiments, R.sup.5 is CH.sub.2CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.5 is C(O)O-methyl. In certain embodiments, R.sup.5 is C(O)O-ethyl. In certain embodiments, R.sup.5 is C(O)O-propyl, OC(O)-isopropyl, C(O)O-isobutyl, or OC(O)-isoamyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.2CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.3CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.4CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.5CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.6CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.5 is (CH.sub.2).sub.sCO.sub.2NR.sup.A.sub.3, wherein R.sup.A is hydrogen or optionally substituted alkyl, and s is 1 to 5.

[0128] In certain embodiments, R.sup.5 is optionally substituted aryl. In some embodiments, R.sup.5 is of the following structure:

##STR00010##

wherein:

[0129] each instance of R.sup.7 is independently selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B;

[0130] each instance of R.sup.A is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl;

[0131] each instance of R.sup.B is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl, or two R.sup.B are taken together with the intervening atoms to form a heterocycle; and

[0132] n is 0, 1, 2, 3, 4 or 5.

[0133] As generally defined herein, n is 0, 1, 2, 3, 4, or 5. In certain embodiments, n is 0 and R.sup.5 is phenyl. In certain embodiments, n is 1 and R.sup.5 is of the formula

##STR00011##

In certain embodiments, n is 1 and R.sup.5 is of the formula

##STR00012##

In certain embodiments, n is 1 and R.sup.5 is of the formula

##STR00013##

In certain embodiments, n is 2 and R.sup.5 is of the formula

##STR00014##

In certain embodiments, n is 2 and R.sup.5 is of the formula

##STR00015##

In certain embodiments, n is 2 and R.sup.5 is of the formula

##STR00016##

In certain embodiments, n is 2 and R.sup.5 is of the formula

##STR00017##

In certain embodiments, In certain embodiments, n is 2 and R.sup.5 is of the formula

##STR00018##

In certain embodiments, In certain embodiments, n is 2 and R.sup.5 is of the formula

##STR00019##

In certain embodiments, n is 3 and R.sup.5 is of the formula

##STR00020##

In certain embodiments, n is 3 and R.sup.5 is of the formula

##STR00021##

In certain embodiments, n is 3 and R.sup.5 is of the formula

##STR00022##

In certain embodiments, n is 3 and R.sup.5 is of the formula

##STR00023##

In certain embodiments, n is 3 and R.sup.5 is of the formula

##STR00024##

In certain embodiments, n is 4 and R.sup.5 is of the formula

##STR00025##

In certain embodiments, n is 4 and R.sup.5 is of the formula

##STR00026##

In certain embodiments, n is 4 and R.sup.5 is of the formula

##STR00027##

In certain embodiments, n is 5 and R.sup.5 is of the formula

##STR00028##

[0134] In certain embodiments, R.sup.7 is hydrogen. In some embodiments, R.sup.7 is halo. In certain embodiments, R.sup.7 is fluoro. In certain embodiments, R.sup.7 is chloro. In certain embodiments, R.sup.7 is bromo. In certain embodiments, R.sup.7 is iodo. In some embodiments, R.sup.7 is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl or optionally substituted aryl. In certain embodiments, R.sup.7 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.7 is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.7 is substituted methyl, ethyl, propyl, butyl, pentyl, or hexyl. In certain embodiments, R.sup.7 is CH.sub.2F. In certain embodiments, R.sup.7 is CHF.sub.2. In certain embodiments, R.sup.7 is CF.sub.3. In certain embodiments, R.sup.7 is (CH.sub.2).sub.sCO.sub.2R.sup.A, wherein s is 1, 2, 3, 4, 5, or 6. In certain embodiments, R.sup.7 is (CH.sub.2).sub.sCO.sub.2H. In some embodiments, s is 1. In some embodiments, s is 2. In some embodiments, s is 3. In some embodiments, s is 4. In some embodiments, s is 5. In some embodiments, s is 6. In certain embodiments, R.sup.7 is CH.sub.2CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is C(O)O-methyl. In certain embodiments, R.sup.7 is C(O)O-ethyl. In certain embodiments, R.sup.7 is C(O)O-propyl, OC(O)-isopropyl, C(O)O-isobutyl, or OC(O)-isoamyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.2CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.3CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.4CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.5CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.6CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl.

[0135] In certain embodiments, R.sup.7 is unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.7 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. In certain embodiments, R.sup.7 is isopropyl, isobutyl, or isoamyl. In certain embodiments, R.sup.7 is isobutyl. In certain embodiments, R.sup.7 is tert-butyl.

[0136] In certain embodiments, R.sup.7 is OR.sup.A. In certain embodiments, R.sup.7 is OH. In certain embodiments, R.sup.7 is OR.sup.A, wherein R.sup.A is optionally substituted alkyl. In certain embodiments, R.sup.7 is O-methyl. In certain embodiments, R.sup.7 is O-ethyl. In certain embodiments, R.sup.7 is O-propyl, O-isopropyl, O-isobutyl, or O-isoamyl.

[0137] In certain embodiments, R.sup.7 is N(R.sup.B).sub.2, wherein each R.sup.B is independently selected from the group consisting of methyl, ethyl, isopropyl, isobutyl, isoamyl, and benzyl. In some embodiments, R.sup.7 is N(R.sup.B).sub.2, wherein each R.sup.B is the same. In some embodiments, R.sup.7 is N(R.sup.B).sub.2, wherein each R.sup.B is different. In certain embodiments, R.sup.7 is NH.sub.2.

[0138] In certain embodiments, R.sup.7 is CO.sub.2R.sup.A. In certain embodiments, R.sup.7 is CO.sub.2H. In certain embodiments, R.sup.7 is CO.sub.2R.sup.A, wherein R.sup.A is optionally substituted alkyl. In certain embodiments, R.sup.7 is C(O)O-methyl. In certain embodiments, R.sup.7 is C(O)O-ethyl. In certain embodiments, R.sup.7 is C(O)O-propyl, OC(O)-isopropyl, C(O)O-isobutyl, or OC(O)-isoamyl.

[0139] In some embodiments, at least two of R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are hydrogen. In certain embodiments, R.sup.1 and R.sup.4 are hydrogen. In certain embodiments, R.sup.3 and R.sup.4 are hydrogen. In certain embodiments, R.sup.1 and R.sup.2 are hydrogen. In certain embodiments, R.sup.1 and R.sup.3 are hydrogen. In certain embodiments, R.sup.2 and R.sup.4 are hydrogen. In some embodiments, at least two of R.sup.1, R.sup.2, R.sup.3, and R.sup.4 are each hydrogen, and R.sup.5 is hydrogen. In some embodiments, R.sup.1, R.sup.3, and R.sup.4 are hydrogen. In some embodiments, R.sup.1, R.sup.2, and R.sup.3 are hydrogen. In some embodiments, R.sup.2, R.sup.3, and R.sup.4 are hydrogen.

[0140] In some embodiments, the compound provided herein is of Formula (III) or (IV):

##STR00029##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.1, R.sup.2, R.sup.3, and R.sup.5 are as defined herein. In certain embodiments, R.sup.2 and R.sup.3 are taken together with their intervening atoms to form an optionally substituted carbocycle or heterocycle.

[0141] In certain embodiments, the compound provided herein is of Formula (III-a) or (III-b):

##STR00030##

[0142] or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.1, R.sup.2, and R.sup.5 are as defined herein. In certain embodiments, a compound of the present disclosure is selected from the group consisting of the compounds listed in Tables 1a and 1b.

[0143] Compounds of interest also include 2,3-dihydro-1H-benzo[de]isoquinoline analogues, e.g., compounds that include a 2,3-dihydro-1H-benzo[de]isoquinoline scaffold substituted with one or more substituents which are non-hydrogen. The 2,3-dihydro-1H-benzo[de]isoquinoline scaffold of Formula (V) may be substituted at any position 1, 2, 3, 4, 5, 6, 7, 8 and/or 9:

##STR00031##

[0144] In certain embodiments, positions 3, 7, 8, and/or 9 of the Formula (V) scaffold are substituted as described herein. In certain embodiments, only positions 3 and 7 of the Formula (V) scaffold are substituted as described herein. In certain embodiments, positions 7, 8, and/or 9 of the Formula (V) scaffold are substituted as described herein. In certain embodiments, positions 7 and 8 of the Formula (V) scaffold are substituted as described herein. In certain embodiments, positions 7 and 9 of the Formula (V) scaffold are substituted as described herein. In certain embodiments, positions 7 of the Formula (V) scaffold are substituted as described herein. In certain embodiments, positions 3, 4, 7, 8, and/or 9 of the Formula (V) scaffold are substituted as described herein. In certain embodiments, positions 4, 7, 8, and 9 of the Formula (V) scaffold are substituted as described herein. In certain embodiments, positions 4 and 7 of the Formula (V) scaffold are substituted as described herein. The remaining positions of the foregoing embodiments are substituted with hydrogen.

TABLE-US-00001 TABLE 1a Exemplary Compounds of Formula I-a. BO globin num- IC.sub.50 induction Reference Yield ID ber Structure (M) (at IC.sub.50) MF MW MP ( C.) MP ( C.) (%) A1 BO- 2388 [00032] >100 ND C.sub.18H.sub.11NO.sub.2 273.3 218-220 202 (16) 70.7 A2 BO- 2375 [00033] 0.3 1.1 C.sub.18H.sub.10N.sub.2O.sub.4 318.3 >280 280-282 (17) 92.7 A3 BO- 2376 [00034] 5.2 ND C.sub.18H.sub.8Cl.sub.2N.sub.2O.sub.4 387.2 >280 85.2 A4 BO- 2377 [00035] 21 ND C.sub.19H.sub.12N.sub.2O.sub.5 348.3 >280 92.2 A5 BO- 2378 [00036] 6.7 ND C.sub.22H.sub.12N.sub.2O.sub.6 404.4 254.5-256 38 A6 BO- 2379 [00037] >100 ND C.sub.18H.sub.10ClNO.sub.2 307.7 263-265.5 231-233 (18) 22.4 A7 BO- 2380 [00038] >100 ND C.sub.19H.sub.12ClNO.sub.3 337.8 >280 37 A8 BO- 2381 [00039] >100 ND C.sub.19H.sub.9ClF.sub.3NO.sub.2 375.7 278-279 17.3 A9 BO- 2382 [00040] >100 ND C.sub.18H.sub.9BrClNO.sub.2 386.6 >280 13.2 A10 BO- 2383 [00041] >100 ND C.sub.18H.sub.9Cl.sub.2NO.sub.2 342.2 273-275 12.1 A11 BO- 2384 [00042] >100 ND C.sub.18H.sub.8BrClFNO.sub.2 404.6 >280 49.2 A12 BO- 2384 [00043] >100 ND C.sub.18H.sub.9ClFNO.sub.2 325.7 274-276 40.5 A13 BO- 2386 [00044] >100 ND C.sub.18H.sub.9BrClNO.sub.2 386.6 >280 66.3 A14 BO- 2387 [00045] >100 ND C.sub.18H.sub.8Cl.sub.3NO.sub.2 376.6 >280 >300 (18) 15.9 B1 BO- 2393 [00046] 10.7 4.1 C.sub.16H.sub.17ClN.sub.2O.sub.2 304.8 >280 296-298 (20) 84.5 B2 BO- 2390 [00047] 2.6 ND C.sub.16H.sub.15N.sub.3O.sub.4 313.3 131-133 106-109 (20) 65.7 B3 BO- 2391 [00048] 2.3 ND C.sub.17H.sub.17N.sub.3O.sub.4 327.3 108-110 106-109 (21) 42 B4 BO- 2392 [00049] 5 3.3 C.sub.16H.sub.16Cl.sub.2N.sub.2O.sub.2 339.2 >280 293-295 (22) 81.5 C1 BO- 2400 [00050] >100 ND C.sub.14H.sub.11NO.sub.2 225.2 171-172.5 158 (18) 37.3 C2 BO- 2396 [00051] >100 ND C.sub.13H.sub.8N.sub.2O.sub.4 256.2 209-212.5 208-209 (18) 51.1 C3 BO- 2397 [00052] >100 ND C.sub.14H.sub.10N.sub.2O.sub.4 270.2 191-192.5 187.5-188.5 (18) 34.1 C4 BO- 2398 [00053] >100 ND C.sub.13H.sub.8ClNO.sub.2 245.7 186-188 171-173 (18) 32.6 C5 BO- 2399 [00054] >100 ND C.sub.14H.sub.10ClNO.sub.2 259.7 167-170 165-166 (18) 33.7 D1 BO- 2401 [00055] 6.3 ND C.sub.18H.sub.16N.sub.2O.sub.6 356.3 115-116 97 E1 BO- 2389 [00056] 2.2 1.3 C.sub.18H.sub.12N.sub.2O.sub.2 288.3 276-278 302-304 (23) 56.4 E2 (SS- 2394) BO- 2394 [00057] 2.3 4 C.sub.16H.sub.18ClN.sub.3O.sub.2 319.8 >280 184-185 (20) 77.3 E3 BO- 2395 [00058] 11.2 2.7 C.sub.17H.sub.20ClN.sub.3O.sub.2 333.8 >280 184-185 (21) 42.8

TABLE-US-00002 TABLE 1b Additional Exemplary Compounds of Formula I-a. BO number Structure BO-2562 [00059] BO-2559 [00060] BO-2561 [00061]

[0145] In certain embodiments, a substituent may contribute to optical isomerism and/or stereo isomerism of a compound. The compound(s) of the present disclosure provided herein include the neutral form, salts, solvates, hydrates, and prodrug forms of a compound. All such forms are embraced by the present disclosure. Thus the compounds described herein include salts, solvates, hydrates, prodrug, and isomer forms thereof, including the pharmaceutically acceptable salts, solvates, hydrates, prodrugs, and isomers thereof. In certain embodiments, a compound may be a metabolized into a pharmaceutically active derivative.

[0146] In certain embodiments, the present disclosure employs a compound of Formula (V-a):

##STR00062##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein:

[0147] R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12 and R.sup.13 are each independently selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B;

[0148] R.sup.14 is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, or optionally substituted aryl;

[0149] R.sup.15 and R.sup.16 are each independently selected from the group consisting of hydrogen, oxygen, hydroxyl, OR, and OC(O)R, wherein both R.sup.15 and R.sup.16 are not oxygen connected via a double bond, wherein R is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl;

[0150] each instance of R.sup.A is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl; and

[0151] each instance of R.sup.B is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl, or two R.sup.B are taken together with the intervening atoms to form a heterocycle.

[0152] In some embodiments, R.sup.10 is N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.A is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.A is independently hydrogen, methyl or ethyl. In some embodiments, R.sup.B is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.B is independently hydrogen, methyl or ethyl. In some embodiments, each R.sup.B is hydrogen. In some embodiments, each R.sup.B is methyl. In some embodiments, R.sup.10 is hydrogen, NH.sub.2, NHC(O)CH.sub.3, NHC(O)OR.sup.A, NHR.sup.B, NR.sup.B.sub.2, wherein R.sup.B can be C.sub.1-C.sub.5 alkyl. In some embodiments, R.sup.10 is hydrogen, NHC(O)CH.sub.3, NHC(O)O(CH.sub.3), N(CH.sub.3).sub.2, NH.sub.2, or NHCH.sub.2CH.sub.3. In some embodiments, R.sup.10 is hydrogen and R.sup.8, R.sup.9, R.sup.11, R.sup.12, and R.sup.13 are each independently H or the groups recited for R.sup.10.

[0153] In some embodiments, R.sup.11 is N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.A is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.A is independently hydrogen, methyl or ethyl. In some embodiments, R.sup.B is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.B is independently hydrogen, methyl or ethyl. In some embodiments, each R.sup.B is hydrogen. In some embodiments, each R.sup.B is methyl. In some embodiments, R.sup.11 is hydrogen, NH.sub.2, NHC(O)CH.sub.3, N(R.sup.A)C(O)OR.sup.B, NHR.sup.B, NR.sup.B.sub.2, wherein R.sup.B can be C.sub.1-C.sub.5 alkyl. In some embodiments, R.sup.11 is hydrogen, NHC(O)CH.sub.3, NHC(O)O(CH.sub.3), N(CH.sub.3).sub.2, NH.sub.2, or NHCH.sub.2CH.sub.3. In some embodiments, R.sup.11 is hydrogen and R.sup.8, R.sup.9, R.sup.11, R.sup.12, and R.sup.13 are each independently H or the groups recited for R.sup.11.

[0154] In certain embodiments, R.sup.14 is hydrogen. In some embodiments, R.sup.14 is an optionally substituted alkyl. In certain embodiments, R.sup.14 is an optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is a substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is a unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is a substituted methyl, ethyl, propyl, butyl, pentyl, or hexyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 1, 2, 3, 4, 5, or 6, and wherein R.sup.B is independently optionally substituted C.sub.1-6 alkyl or two R.sup.B are taken together with the intervening atoms to form a heterocycle with at least 1-4 heteroatoms. In certain embodiments, the heterocycle is a 3-8 membered ring. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 1, 2, 3, 4, or 5, and wherein R.sup.B is independently optionally substituted C.sub.1-6 alkyl or two R.sup.B are taken together with the intervening atoms to form a heterocycle. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, n is 4. In certain embodiments, n is 5. In certain embodiments, n is 6. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 2. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is unsubstituted C.sub.1-6 alkyl. In certain embodiment, R.sup.14 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.3)R.sup.B, wherein each R.sup.B is independently optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.3)R.sup.B, wherein each R.sup.B is independently unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.3).sub.2. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.2CH.sub.3)R.sup.B, wherein each R.sup.B is independently optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.2CH.sub.3)R.sup.B, wherein each R.sup.B is independently unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is independently selected from the group consisting of methyl, ethyl, isopropyl, isobutyl, isoamyl, and benzyl. In some embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is the same. In some embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is different. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nNH.sub.2. In certain embodiments, R.sup.14 is (CH.sub.2).sub.2N(CH.sub.3).sub.2. In certain embodiments, R.sup.14 comprises a carboxylic acid or tetra-ammonium moiety. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nCOOH or (CH.sub.2).sub.nCOONR.sup.C.sub.3 or, wherein n is 1, 2, 3, 4, 5, or 6 and R.sup.C is an optionally substituted C.sub.1-6 alkyl, wherein the optional substituent is a hydroxyl. In certain embodiments, R.sup.C is (CH.sub.2).sub.2OH, (CH.sub.2).sub.2OH, (CH.sub.2).sub.3OH, (CH.sub.2).sub.4OH, (CH.sub.2).sub.5OH.

[0155] As described generally above, R.sup.8 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.8 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.8 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.8 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.8 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.8 is hydrogen. In some embodiments, R.sup.8 is C.sub.1-6 alkyl. In certain embodiments, R.sup.8 is C(O)OR.sup.A. In certain embodiments, R.sup.8 is CO.sub.2H. In certain embodiments, R.sup.8 is C(O)N(R.sup.B).sub.2 or N(R.sup.A)C(O)R.sup.A. In some embodiments, R.sup.8 is halogen. In certain embodiments, R.sup.8 is fluoro. In certain embodiments, R.sup.8 is chloro. In certain embodiments, R.sup.8 is bromo. In certain embodiments, R.sup.8 is iodo. In some embodiments, R.sup.8 is NH.sub.2. In some embodiments, R.sup.8 is N(R.sup.B).sub.2. In some embodiments, R.sup.8 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.8 is NHCH.sub.3. In certain embodiments, R.sup.8 is NO.sub.2. In some embodiments, R.sup.8 is optionally substituted naphthyl.

[0156] As described generally above, R.sup.9 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.9 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.9 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.9 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.9 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.9 is hydrogen. In some embodiments, R.sup.9 is C.sub.1-6 alkyl. In certain embodiments, R.sup.9 is C(O)OR.sup.A. In certain embodiments, R.sup.9 is CO.sub.2H. In certain embodiments, R.sup.9 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.9 is halogen. In certain embodiments, R.sup.9 is fluoro. In certain embodiments, R.sup.9 is chloro. In certain embodiments, R.sup.9 is bromo. In certain embodiments, R.sup.9 is iodo. In some embodiments, R.sup.9 is N(R.sup.B).sub.2. In some embodiments, R.sup.9 is NH.sub.2. In some embodiments, R.sup.9 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.9 is NHCH.sub.3. In certain embodiments, R.sup.9 is NO.sub.2. In some embodiments, R.sup.9 is optionally substituted naphthyl.

[0157] As described generally above, R.sup.10 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.10 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.10 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.10 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.10 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.10 is hydrogen. In some embodiments, R.sup.10 is C.sub.1-6 alkyl. In certain embodiments, R.sup.10 is C(O)OR.sup.A. In certain embodiments, R.sup.10 is CO.sub.2H. In certain embodiments, R.sup.10 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.10 is halogen. In certain embodiments, R.sup.10 is fluoro. In certain embodiments, R.sup.10 is chloro. In certain embodiments, R.sup.10 is bromo. In certain embodiments, R.sup.10 is iodo. In some embodiments, R.sup.10 is N(R.sup.B).sub.2. In some embodiments, R.sup.10 is NH.sub.2. In some embodiments, R.sup.10 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.10 is NHCH.sub.3. In certain embodiments, R.sup.10 is NO.sub.2. In some embodiments, R.sup.10 is optionally substituted naphthyl. In some embodiments, R.sup.10 is N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.A is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.A is independently hydrogen, methyl or ethyl. In some embodiments, R.sup.B is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.B is independently hydrogen, methyl or ethyl. In some embodiments, each R.sup.B is hydrogen. In some embodiments, each R.sup.B is methyl. In some embodiments, R.sup.10 is hydrogen, NH.sub.2, NHC(O)CH.sub.3, NHC(O)O(R.sup.A), NHR.sup.B, NR.sup.B.sub.2, wherein R.sup.B can be C.sub.1-C.sub.5 alkyl. In some embodiments, R.sup.10 is hydrogen, NHC(O)CH.sub.3, NHC(O)O(CH.sub.3), N(CH.sub.3).sub.2, NH.sub.2, or NHCH.sub.2CH.sub.3.

[0158] As described generally above, R.sup.11 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.11 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.11 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.11 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.11 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.11 is hydrogen. In some embodiments, R.sup.11 is C.sub.1-6 alkyl. In certain embodiments, R.sup.11 is C(O)OR.sup.A. In certain embodiments, R.sup.11 is CO.sub.2H. In certain embodiments, R.sup.11 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.11 is halogen. In certain embodiments, R.sup.11 is fluoro. In certain embodiments, R.sup.11 is chloro. In certain embodiments, R.sup.11 is bromo. In certain embodiments, R.sup.11 is iodo. In some embodiments, R.sup.11 is N(R.sup.B).sub.2. In some embodiments, R.sup.11 is NH.sub.2. In some embodiments, R.sup.11 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.11 is NHCH.sub.3. In certain embodiments, R.sup.11 is NO.sub.2. In some embodiments, R.sup.11 is optionally substituted naphthyl. In some embodiments, R.sup.11 is N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.A is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.A is independently hydrogen, methyl or ethyl. In some embodiments, R.sup.B is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.B is independently hydrogen, methyl or ethyl. In some embodiments, each R.sup.B is hydrogen. In some embodiments, each R.sup.B is methyl. In some embodiments, R.sup.11 is hydrogen, NH.sub.2, NHC(O)CH.sub.3, NHC(O)O(R.sup.A), NHR.sup.B, NR.sup.B.sub.2, wherein R.sup.B can be C.sub.1-C.sub.5 alkyl. In some embodiments, R.sup.11 is hydrogen, NHC(O)CH.sub.3, NHC(O)O(CH.sub.3), N(CH.sub.3).sub.2, NH.sub.2, or NHCH.sub.2CH.sub.3.

[0159] As described generally above, R.sup.12 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.AC(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.12 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.12 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.12 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.12 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.12 is hydrogen. In some embodiments, R.sup.12 is C.sub.1-6 alkyl. In certain embodiments, R.sup.12 is C(O)OR.sup.A. In certain embodiments, R.sup.12 is CO.sub.2H. In certain embodiments, R.sup.12 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.12 is halogen. In certain embodiments, R.sup.12 is fluoro. In certain embodiments, R.sup.12 is chloro. In certain embodiments, R.sup.12 is bromo. In certain embodiments, R.sup.12 is iodo. In some embodiments, R.sup.12 is N(R.sup.B).sub.2. In some embodiments, R.sup.12 is NH.sub.2. In some embodiments, R.sup.12 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.12 is NHCH.sub.3. In certain embodiments, R.sup.12 is NO.sub.2. In some embodiments, R.sup.12 is optionally substituted naphthyl.

[0160] As described generally above, R.sup.13 is selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.13 is selected from the group consisting of halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B. In certain embodiments, R.sup.13 is selected from the group consisting of H, OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.13 is selected from the group consisting of OH, Cl, Br, F, optionally substituted C.sub.1-6 alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted aryl, OR.sup.A, NO.sub.2, N(R.sup.B).sub.2, C(O)CH.sub.3, CO.sub.2H, C(O)OR.sup.A, C(O)N(R.sup.B).sub.2, CN, heterocyclyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.13 is selected from the group consisting of H, OH, Cl, Br, F, methyl, ethyl, methoxy, ethoxy, CC-aryl, phenyl, naphthyl, NO.sub.2, NHC.sub.1-6 alkyl, C(O)CH.sub.3, CO.sub.2H, CO.sub.2Et, CONH-aryl, CN, N-morpholinyl, SO.sub.2-alkyl, and SO.sub.2-aryl. In certain embodiments, R.sup.13 is hydrogen. In some embodiments, R.sup.13 is C.sub.1-6 alkyl. In certain embodiments, R.sup.13 is C(O)OR.sup.A. In certain embodiments, R.sup.13 is CO.sub.2H. In certain embodiments, R.sup.13 is C(O)N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A or N(R.sup.A)C(O)OR.sup.A. In some embodiments, R.sup.13 is halogen. In certain embodiments, R.sup.13 is fluoro. In certain embodiments, R.sup.13 is chloro. In certain embodiments, R.sup.13 is bromo. In certain embodiments, R.sup.13 is iodo. In some embodiments, R.sup.13 is N(R.sup.B).sub.2. In some embodiments, R.sup.13 is NH.sub.2. In some embodiments, R.sup.13 is NHC.sub.1-6 alkyl. In certain embodiments, R.sup.13 is NHCH.sub.3. In certain embodiments, R.sup.13 is NO.sub.2. In some embodiments, R.sup.13 is optionally substituted naphthyl.

[0161] As described generally above, R.sup.14 is hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, or optionally substituted aryl. In certain embodiments, R.sup.14 is hydrogen. In some embodiments, R.sup.14 is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl or optionally substituted aryl. In certain embodiments, R.sup.14 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. In certain embodiments, R.sup.14 is isopropyl, isobutyl, or isoamyl. In certain embodiments, R.sup.14 is isobutyl. In certain embodiments, R.sup.14 is tert-butyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 1, 2, 3, 4, 5, or 6 and wherein R.sup.B is independently optionally substituted C.sub.1-6 alkyl or two R.sup.B are taken together with the intervening atoms to form a heterocycle with at least 1-4 heteroatoms. In certain embodiments, the heterocycle is a 3-8 membered ring. In certain embodiments, the heterocycle formed is In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 1, 2, 3, 4, or 5, and wherein R.sup.B is independently optionally substituted C.sub.1-6 alkyl or two R.sup.B are taken together with the intervening atoms to form a heterocycle. In certain embodiments, n is 1. In certain embodiments, n is 2. In certain embodiments, n is 3. In certain embodiments, n is 4. In certain embodiments, n is 5. In certain embodiments, n is 6. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nNHR.sup.B. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is unsubstituted C.sub.1-6 alkyl. In certain embodiment, R.sup.14 is (CH.sub.2).sub.nNHR.sup.B, wherein R.sup.B is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.3)R.sup.B, wherein each R.sup.B is independently optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.3)R.sup.B, wherein each R.sup.B is independently unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.3).sub.2. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.2CH.sub.3)R.sup.B, wherein each R.sup.B is independently optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(CH.sub.2CH.sub.3)R.sup.B, wherein each R.sup.B is independently unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is independently selected from the group consisting of methyl, ethyl, isopropyl, isobutyl, isoamyl, and benzyl. In some embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is the same. In some embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein each R.sup.B is different. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nNH.sub.2. In certain embodiments, R.sup.14 is (CH.sub.2).sub.2N(CH.sub.3).sub.2. In certain embodiments, R.sup.14 comprises a carboxylic acid or tetra-ammonium moiety. In certain embodiments, R.sup.14 is (CH.sub.2).sub.sCO.sub.2R.sup.A, wherein s is 1, 2, 3, 4, 5, or 6. In certain embodiments, R.sup.14 is (CH.sub.2).sub.sCO.sub.2H. In some embodiments, s is 1. In some embodiments, s is 2. In some embodiments, s is 3. In some embodiments, s is 4. In some embodiments, s is 5. In some embodiments, s is 6. In certain embodiments, R.sup.14 is CH.sub.2CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.14 is CH.sub.2CO.sub.2NR.sup.c.sub.3, wherein R.sup.C is an optionally substituted C.sub.1-6 alkyl, wherein the optional substituent is a hydroxyl. In certain embodiments, R.sup.C is (CH.sub.2).sub.2OH, (CH.sub.2).sub.2OH, (CH.sub.2).sub.3OH, (CH.sub.2).sub.4OH, (CH.sub.2).sub.5OH. In certain embodiments, R.sup.14 is C(O)O-methyl. In certain embodiments, R.sup.14 is C(O)O-ethyl. In certain embodiments, R.sup.14 is C(O)O-propyl, OC(O)-isopropyl, C(O)O-isobutyl, or OC(O)-isoamyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.2CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.3CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.4CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.5CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.6CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl.

[0162] In certain embodiments, R.sup.14 is optionally substituted aryl. In some embodiments, R.sup.14 is of the following structure:

##STR00063##

wherein:

[0163] each instance of R.sup.7 is independently selected from the group consisting of hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, OR.sup.A, OC(O)R.sup.A, SR.sup.A, N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, C(O)N(R.sup.B).sub.2, CN, NO.sub.2, C(O)R.sup.A, C(O)OR.sup.A, S(O)R.sup.A, SO.sub.2R.sup.A, SO.sub.2N(R.sup.B).sub.2, and NHSO.sub.2R.sup.B;

[0164] each instance of R.sup.A is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl;

[0165] each instance of R.sup.B is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl, optionally substituted heteroaryl, and optionally substituted aryl, or two R.sup.B are taken together with the intervening atoms to form a heterocycle; and

[0166] n is 0, 1, 2, 3, 4 or 5.

[0167] As generally defined herein, n is 0, 1, 2, 3, 4, or 5. In certain embodiments, n is 0 and R.sup.14 is phenyl. In certain embodiments, n is 1 and R.sup.14 is of the formula

##STR00064##

In certain embodiments, n is 1 and R.sup.14 is of the formula.

##STR00065##

In certain embodiments, n is 1 and R.sup.14 is of the formula

##STR00066##

certain embodiments, n is 2 and R.sup.14 is of the formula

##STR00067##

In certain embodiments, n is 2 and R.sup.14 is of the formula

##STR00068##

In certain embodiments, n is 2 and R.sup.14 is of the formula

##STR00069##

In certain embodiments, n is 2 and R.sup.14 is of the formula

##STR00070##

In certain embodiments, In certain embodiments, n is 2 and R.sup.14 is of the formula

##STR00071##

In certain embodiments, In certain embodiments, n is 2 and R.sup.14 is of the formula

##STR00072##

In certain embodiments, n is 3 and R.sup.14 is of the formula

##STR00073##

In certain embodiments, n is 3 and R.sup.14 is of the formula

##STR00074##

In certain embodiments, n is 3 and R.sup.14 is of the formula

##STR00075##

In certain embodiments, n is 3 and R.sup.14 is of the formula

##STR00076##

In certain embodiments, n is 3 and R.sup.14 is of the formula

##STR00077##

In certain embodiments, n is 4 and R.sup.14 is of the formula

##STR00078##

In certain embodiments, n is 4 and R.sup.14 is of the formula

##STR00079##

In certain embodiments, n is 4 and R.sup.14 is of the formula

##STR00080##

In certain embodiments, n is 5 and R.sup.14 is of the formula

##STR00081##

[0168] In certain embodiments, R.sup.7 is hydrogen. In some embodiments, R.sup.7 is halo. In certain embodiments, R.sup.7 is fluoro. In certain embodiments, R.sup.7 is chloro. In certain embodiments, R.sup.7 is bromo. In certain embodiments, R.sup.7 is iodo. In some embodiments, R.sup.7 is optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heterocyclyl or optionally substituted aryl. In certain embodiments, R.sup.7 is optionally substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.7 is substituted C.sub.1-6 alkyl. In certain embodiments, R.sup.7 is substituted methyl, ethyl, propyl, butyl, pentyl, or hexyl. In certain embodiments, R.sup.7 is CH.sub.2F. In certain embodiments, R.sup.7 is CHF.sub.2. In certain embodiments, R.sup.7 is CF.sub.3. In certain embodiments, R.sup.7 is (CH.sub.2).sub.sCO.sub.2R.sup.A, wherein s is 1, 2, 3, 4, 5, or 6. In certain embodiments, R.sup.7 is (CH.sub.2).sub.sCO.sub.2H. In some embodiments, s is 1. In some embodiments, s is 2. In some embodiments, s is 3. In some embodiments, s is 4. In some embodiments, s is 5. In some embodiments, s is 6. In certain embodiments, R.sup.7 is CH.sub.2CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is C(O)O-methyl. In certain embodiments, R.sup.7 is C(O)O-ethyl. In certain embodiments, R.sup.7 is C(O)O-propyl, OC(O)-isopropyl, C(O)O-isobutyl, or OC(O)-isoamyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.2CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.3CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.4CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.5CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl. In certain embodiments, R.sup.7 is (CH.sub.2).sub.6CO.sub.2R.sup.A, wherein R.sup.A is hydrogen or optionally substituted alkyl.

[0169] In certain embodiments, R.sup.7 is unsubstituted C.sub.1-6 alkyl. In certain embodiments, R.sup.7 is methyl, ethyl, propyl, butyl, pentyl, or hexyl. In certain embodiments, R.sup.7 is isopropyl, isobutyl, or isoamyl. In certain embodiments, R.sup.7 is isobutyl. In certain embodiments, R.sup.7 is tert-butyl.

[0170] In certain embodiments, R.sup.7 is OR.sup.A. In certain embodiments, R.sup.7 is OH. In certain embodiments, R.sup.7 is OR.sup.A, wherein R.sup.A is optionally substituted alkyl. In certain embodiments, R.sup.7 is O-methyl. In certain embodiments, R.sup.7 is O-ethyl. In certain embodiments, R.sup.7 is O-propyl, O-isopropyl, O-isobutyl, or O-isoamyl.

[0171] In certain embodiments, R.sup.7 is N(R.sup.B).sub.2, wherein each R.sup.B is independently selected from the group consisting of methyl, ethyl, isopropyl, isobutyl, isoamyl, and benzyl. In some embodiments, R.sup.7 is N(R.sup.B).sub.2, wherein each R.sup.B is the same. In some embodiments, R.sup.7 is N(R.sup.B).sub.2, wherein each R.sup.B is different. In certain embodiments, R.sup.7 is NH.sub.2.

[0172] In certain embodiments, R.sup.7 is CO.sub.2R.sup.A. In certain embodiments, R.sup.7 is CO.sub.2H. In certain embodiments, R.sup.7 is CO.sub.2R.sup.A, wherein R.sup.A is optionally substituted alkyl. In certain embodiments, R.sup.7 is C(O)O-methyl. In certain embodiments, R.sup.7 is C(O)O-ethyl. In certain embodiments, R.sup.7 is C(O)O-propyl, OC(O)-isopropyl, C(O)O-isobutyl, or OC(O)-isoamyl.

[0173] In some embodiments, at least two of R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, and R.sup.13 are hydrogen. In certain embodiments, R.sup.8, R.sup.9, R.sup.12, and R.sup.13 are hydrogen. In certain embodiments, R.sup.12, and R.sup.13 are are hydrogen. In certain embodiments, R.sup.8, R.sup.9, R.sup.10 are hydrogen. In certain embodiments, R.sup.8, R.sup.9, and R.sup.11 are hydrogen. In certain embodiments, R.sup.10 and R.sup.12 are hydrogen. In some embodiments, at least two of R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, and R.sup.13 are each hydrogen, and R.sup.14 is hydrogen. In some embodiments, R.sup.8, R.sup.9, R.sup.11, R.sup.12, and R.sup.13 are hydrogen.

[0174] In some embodiments of Formula (V-a), R.sup.15 and R.sup.16 are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (VI):

##STR00082##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, and R.sup.14 are as described herein for Formula (V-a).

[0175] In some embodiments of Formula (VI), R.sup.8, R.sup.9, R.sup.11, R.sup.12, and R.sup.13 are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (VI-a):

##STR00083##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.10 and R.sup.14 are as described herein for Formula (V-a). In some examples, R.sup.10 and R.sup.14, independently, are hydrogen, optionally substituted C.sub.1-6 alkyl, NR.sup.B.sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A or hydroxyl.

[0176] In some embodiments of Formula (V-a), R.sup.15 is hydroxyl and R.sup.16 is oxygen connected via a double bond. In certain embodiments, the present disclosure employs a compound of Formula (VII):

##STR00084##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, and R.sup.14 are as described herein for Formula (V-a).

[0177] In some embodiments of Formula (VII), R.sup.8, R.sup.9, R.sup.11, R.sup.12, and R.sup.13 are each hydrogen. In some embodiments, R.sup.8, R.sup.9, R.sup.10, R.sup.12, and R.sup.13 are each hydrogen. In some embodiments, R.sup.8, R.sup.9, R.sup.12, and R.sup.13 are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (VII-a):

##STR00085##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.10, R.sup.11, and R.sup.14 are as described herein for Formula (V-a). In some examples, R.sup.10, R.sup.11, and R.sup.14 independently, are hydrogen, optionally substituted C.sub.1-6 alkyl, NR.sup.B.sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A or hydroxyl.

[0178] In some embodiments of Formula (V-a), R.sup.15 is hydrogen and R.sup.16 is oxygen connected via a double bond. In certain embodiments, the present disclosure employs a compound of Formula (VIII):

##STR00086##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, R.sup.14 are as described herein for Formula (V-a).

[0179] In some embodiments of Formula (VIII), R.sup.8, R.sup.9, R.sup.11, R.sup.12, R.sup.13 are each hydrogen. In some embodiments, R.sup.8, R.sup.9, R.sup.10, R.sup.12, R.sup.13, and R.sup.15 are each hydrogen. In some embodiments, R.sup.8, R.sup.9, R.sup.12, R.sup.13, and R.sup.15 are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (VIII-a):

##STR00087##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.10, R.sup.11, and R.sup.14 are as described herein for Formula (V-a). In some examples, R.sup.10, R.sup.11, and R.sup.14 independently, are hydrogen, optionally substituted C.sub.1-6 alkyl, NR.sup.B.sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A or hydroxyl.

[0180] In some embodiments of Formula (V-a), R.sup.15 is hydrogen and R.sup.16 is hydroxyl. In certain embodiments, the present disclosure employs a compound of Formula (IX):

##STR00088##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13 and R.sup.14 are as described herein for Formula (V-a).

[0181] In some embodiments of Formula (IX), R.sup.8, R.sup.9, R.sup.12, R.sup.13, and R.sup.15 are each hydrogen. In some embodiments of Formula (IX), R.sup.8, R.sup.9, R.sup.10, R.sup.12, R.sup.13, and R.sup.15 are each hydrogen. In some embodiments of Formula (IX), R.sup.8, R.sup.9, R.sup.11, R.sup.12, R.sup.13, and R.sup.15 are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (IX-a):

##STR00089##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.10, R.sup.11, and R.sup.14 are as described herein for Formula (V-a). In some examples, R.sup.10, R.sup.11, and R.sup.14 independently, are hydrogen, optionally substituted C.sub.1-6 alkyl, NR.sup.B.sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A or hydroxyl.

[0182] In some embodiments of Formula (V-a), R.sup.15 and R.sup.16 are each hydroxyl. In certain embodiments, the present disclosure employs a compound of Formula (X):

##STR00090##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13 and R.sup.14 are as described herein for Formula (V-a).

[0183] In some embodiments, R.sup.8, R.sup.9, R.sup.11, R.sup.12, and R.sup.13 of Formula (X) are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (X):

##STR00091##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.10 and R.sup.14 are as described herein for Formula (V-a). In some examples, R.sup.10 and R.sup.14, independently, are hydrogen, optionally substituted C.sub.1-6 alkyl, NR.sup.B.sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A or hydroxyl.

[0184] In some embodiments of Formula (V-a), R.sup.15 and R.sup.16 are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (XI):

##STR00092##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.8, R.sup.9, R.sup.10, R.sup.11, R.sup.12, R.sup.13, and R.sup.14 are as described herein for Formula (V-a).

[0185] In some embodiments, R.sup.8, R.sup.9, R.sup.11, R.sup.12, and R.sup.13 of Formula (XI) are each hydrogen. In certain embodiments, the present disclosure employs a compound of Formula (XI-a):

##STR00093##

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R.sup.10 and R.sup.14 are as described herein for Formula (V-a). In some examples, R.sup.10 and R.sup.14, independently, are hydrogen, optionally substituted C.sub.1-6 alkyl, NR.sup.B.sub.2, N(R.sup.A)C(O)R.sup.A, N(R.sup.A)C(O)OR.sup.A or hydroxyl.

[0186] In some embodiments of the present disclosure is a compound represented by formula (V-a), (VI), (VI-a), (VII), (VII-a), (VIII), (VIII-a), (IX), (IX-a), (X), (X-a), (XI), and/or (XI-a) or a pharmaceutically acceptable salt thereof, wherein R.sup.10 is N(R.sup.B).sub.2, N(R.sup.A)C(O)R.sup.A, or N(R.sup.A)C(O)OR.sup.A and R.sup.14 is an optionally substituted alkyl. In some embodiments, R.sup.A is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.A is independently hydrogen, methyl or ethyl. In some embodiments, R.sup.B is independently hydrogen or optionally substituted alkyl. In some embodiments, R.sup.B is independently hydrogen, methyl or ethyl. In some embodiments, each R.sup.B is hydrogen. In some embodiments, each R.sup.B is methyl. In some embodiments, R.sup.10 is hydrogen, NH.sub.2, NHC(O)CH.sub.3, NHC(O)OR.sup.A, NHR.sup.B, NR.sup.B.sub.2, wherein R.sup.B can be C.sub.1-C.sub.5 alkyl. In some embodiments, R.sup.10 is hydrogen, NHC(O)CH.sub.3, NHC(O)O(CH.sub.3), N(CH.sub.3).sub.2, NH.sub.2, or NHCH.sub.2CH.sub.3. In some embodiments, R.sup.14 is an optionally substituted alkyl. In certain embodiments, R.sup.14 is an optionally substituted methyl, ethyl, propyl, butyl, pentyl, or hexyl. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 1, 2, 3, 4, 5, or 6 and wherein R.sup.B is independently optionally substituted C.sub.1-6 alkyl or two R.sup.B are taken together with the intervening atoms to form a heterocycle with at least 1-4 heteroatoms. In certain embodiments, the heterocycle is a 3-8 membered ring. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nN(R.sup.B).sub.2, wherein n is 2. In certain embodiments, R.sup.14 is (CH.sub.2).sub.2NH.sub.2. In certain embodiments, R.sup.14 comprises a carboxylic acid or tetra-ammonium moiety. In certain embodiments, R.sup.14 is (CH.sub.2).sub.nCOOH or (CH.sub.2).sub.nCOONR.sup.C.sub.3 or, wherein n is 1, 2, 3, 4, 5, or 6 and R.sup.C is an optionally substituted C.sub.1-6 alkyl, wherein the optional substituent is a hydroxyl. In certain embodiments, R.sup.C is (CH.sub.2).sub.2OH, (CH.sub.2).sub.2OH, (CH.sub.2).sub.3OH, (CH.sub.2).sub.4OH, (CH.sub.2).sub.5OH.

[0187] In certain embodiments, exemplary compounds of the present disclosure is one of the compounds provided below or analogues thereof:

##STR00094##

[0188] In some embodiments, a compound of the present disclosure is 1H-benzo[de]isoquinoline-1,3(2H)-dione or analogues thereof as represented by formula A, I-a, II, III, IV, III-a, or III-b. In some embodiments, a compound of the present disclosure is 2,3-dihydro-1H-benzo[de]isoquinoline-1-one or analogues thereof as represented by formula V-a, VIII, or VIII-a. In some embodiments, a compound of the present disclosure is 2,3-dihydro-1H-benzo[de]isoquinoline or analogues thereof as represented by formula V-a, XI, or XI-a. In some embodiments, a compound of the present disclosure is 2,3-dihydro-1H-benzo[de]isoquinoline-1,3-diol or analogues thereof as represented by formula V, X, or X-a. In some embodiments, a compound of the present disclosure is 2,3-dihydro-1H-benzo[de]isoquinolin-1-ol or analogues thereof as represented by formula V, IX, or IX-a. In some embodiments, a compound of the present disclosure is 3-hydroxy-2,3-dihydro-1H-benzo[de]isoquinoline-1-one or analogues thereof as represented by formula V, VII, or VII-a.

[0189] In an alternative embodiment, a compound of the present disclosure is depicted by a compound in Table 2 or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

TABLE-US-00003 TABLE 2 Structures of Examplary Compounds BO number Structure BO-2566 [00095] BO-2563 [00096] BO-2560 [00097] BO-2565 [00098] BO-2477 [00099] BO-2476 [00100] BO-2478 [00101]

Pharmaceutical Compositions

[0190] Pharmaceutical compositions of the present disclosure and for use in accordance with the present disclosure may include a pharmaceutically acceptable excipient or carrier. As used herein, the term pharmaceutically acceptable carrier or pharmaceutically acceptable excipient means a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1980) discloses various excipients used in formulating pharmaceutical compositions and known techniques for the preparation thereof. Some examples of materials which can serve as pharmaceutically acceptable carriers are sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; detergents such as Tween 80; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; and phosphate buffer solutions, as well as other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the composition, according to the judgment of the formulator. The pharmaceutical compositions of this present disclosure can be administered to humans and/or to animals, orally, rectally, parenterally, intracisternally, intravaginally, intranasally, intraperitoneally, topically (as by powders, creams, ointments, or drops), bucally, or as an oral or nasal spray.

[0191] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredients (i.e., microparticles, nanoparticles, liposomes, micelles, polynucleotide/lipid complexes), the liquid dosage forms may contain inert diluents commonly used in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0192] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension, or emulsion in a nontoxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables. In certain embodiments, the particles are suspended in a carrier fluid comprising 1% (w/v) sodium carboxymethyl cellulose and 0.1% (v/v) Tween 80.

[0193] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0194] In order to prolong the effect of a drug, it is often desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension or crystalline or amorphous material with poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions which are compatible with body tissues.

[0195] Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by mixing the particles with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the particles.

[0196] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the particles are mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, c) humectants such as glycerol, d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also comprise buffering agents.

[0197] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols, and the like.

[0198] The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes.

[0199] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.

[0200] Dosage forms for topical or transdermal administration of an inventive pharmaceutical composition include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The particles are admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives or buffers as may be required. Ophthalmic formulation, ear drops, and eye drops are also contemplated as being within the scope of this disclosure.

[0201] In certain embodiments, the pharmaceutically acceptable topical formulations of the present disclosure comprise at least a compound described herein and a penetration enhancing agent. The choice of topical formulation will depend or several factors, including the condition to be treated, the physicochemical characteristics of the inventive compound and other excipients present, their stability in the formulation, available manufacturing equipment, and costs constraints. As used herein the term penetration enhancing agent means an agent capable of transporting a pharmacologically active compound through the stratum coreum and into the epidermis or dermis, preferably, with little or no systemic absorption. A wide variety of compounds have been evaluated as to their effectiveness in enhancing the rate of penetration of drugs through the skin. See, for example, Percutaneous Penetration Enhancers, Maibach H. I. and Smith H. E. (eds.), CRC Press, Inc., Boca Raton, Fla. (1995), which surveys the use and testing of various skin penetration enhancers, and Buyuktimkin et al., Chemical Means of Transdermal Drug Permeation Enhancement in Transdermal and Topical Drug Delivery Systems, Gosh T. K., Pfister W. R., Yum S. I. (Eds.), Interpharm Press Inc., Buffalo Grove, Ill. (1997). In certain exemplary embodiments, penetration agents for use with the present disclosure include, but are not limited to, triglycerides (e.g., soybean oil), aloe compositions (e.g., aloe-vera gel), ethyl alcohol, isopropyl alcohol, octolyphenylpolyethylene glycol, oleic acid, polyethylene glycol 400, propylene glycol, N-decylmethylsulfoxide, fatty acid esters (e.g., isopropyl myristate, methyl laurate, glycerol monooleate, and propylene glycol monooleate), and N-methyl pyrrolidone.

[0202] Transdermal patches have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms can be made by dissolving or dispensing the microparticles or nanoparticles in a proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the particles in a polymer matrix or gel.

[0203] In certain embodiments, the compositions may be in the form of ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants. or patches. In certain exemplary embodiments, formulations of the compositions according to the present disclosure are creams, which may further contain saturated or unsaturated fatty acids such as stearic acid, palmitic acid, oleic acid, palmito-oleic acid, cetyl or oleyl alcohols, stearic acid being particularly preferred. Creams of the present disclosure may also contain a non-ionic surfactant, for example, polyoxy-40-stearate. In certain embodiments, the active component is admixed under sterile conditions with a pharmaceutically acceptable excipient and any needed preservatives or buffers as may be required. Ophthalmic formulations, eardrops, and eye drops are also contemplated as being within the scope of this disclosure. Additionally, the present disclosure contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of a compound to the body. Such dosage forms are made by dissolving or dispensing the compound in the proper medium. As discussed above, penetration enhancing agents can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate controlling membrane or by dispersing the compound in a polymer matrix (e.g., PLGA) or gel.

[0204] The ointments, pastes, creams, and gels may contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc, and zinc oxide, or mixtures thereof.

[0205] Powders and sprays can contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates, and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants such as chlorofluorohydrocarbons.

[0206] It will also be appreciated that the compounds and pharmaceutical compositions of the present disclosure can be formulated and employed in combination therapies, that is, the compounds and pharmaceutical compositions can be formulated with or administered concurrently with, prior to, or subsequent to, one or more other desired therapeutics or medical procedures. The particular combination of therapies (therapeutics or procedures) to employ in a combination regimen will take into account compatibility of the desired therapeutics and/or procedures and the desired therapeutic effect to be achieved. It will also be appreciated that the therapies employed may achieve a desired effect for the same disorder (for example, a provided compound may be administered concurrently with another agent effective against -thalassemia or sickle cell disease), or they may achieve different effects (e.g., control of any adverse effects).

[0207] It will also be appreciated that certain of the compounds of present disclosure can exist in free form for treatment, or where appropriate, as a pharmaceutically acceptable derivative thereof. According to the present disclosure, a pharmaceutically acceptable derivative includes, but is not limited to, pharmaceutically acceptable salts, esters, salts of such esters, or a prodrug or other adduct or derivative of a compound described herein which upon administration to a patient in need is capable of providing, directly or indirectly, a compound as otherwise described herein, or a metabolite or residue thereof.

Methods of Treatment

[0208] The present disclosure provides compounds that induce globin and can produce beneficial therapeutic effects. In certain embodiments, compounds and compositions described herein are used to treat hemoglobinopathies such as sickle cell anemia or -thalassemia. In certain embodiments, a provided compound or composition is used to treat sickle cell anemia. In certain other embodiments, a provided compound or composition is used to treat -thalassemia.

[0209] Without being bound by theory, the compounds (non-HDACi compounds) described herein can stimulate globin expression via regulating both the p38/MAPK signaling pathway and globin gene regulators. As shown in the Examples below, several compounds described herein exhibited better gamma globin-inducing capabilities than hydroxyurea, including higher gamma globin level and superior therapeutic effect (IC.sub.50/EC). Most importantly, those compounds are able to induce globin gene expression in both hydroxyurea-responsive cells and hydroxyurea-resistant cells.

[0210] In one aspect, the present disclosure provides methods comprising the compounds described herein, e.g., compounds of Formula A, I, I-a, II, III, IV, III-a, III-b, V-a, VI, VII, VIII, IX, X or XI, or compositions thereof for stimulating globin expression comprising: contacting a subject with a compound or a composition described herein under conditions suitable to induce globin expression in the subject.

[0211] In certain embodiments, the present disclosure provides a method of inducing globin comprising: contacting a cell with an effective amount of a compound of Formula A, I, I-a, II, III, IV, III-a, III-b, V-a, VI, VII, VIII, IX, X or XI. In certain embodiments, the present disclosure provides a method of inducing globin comprising: administering to a subject an effective amount of a compound of Formula A, I, I-a, II, III, IV, III-a, III-b, V-a, VI, VII, VIII, IX, X or XI.

[0212] In certain embodiments, the present disclosure provides a method of treating 3-thalassemia or sickle cell anemia, the method comprising: administering an effective amount of one or more of the provided compounds or composition to a patient suffering from, suspected of having, or at risk for -thalassemia or sickle cell anemia.

[0213] In certain embodiments, a provided compound or composition is administered orally. In certain embodiments, a provided compound or composition is administered parenterally. In certain embodiments, a provided compound or composition is administered in combination with an additional therapeutic agent.

[0214] The compounds of the present disclosure are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. The expression dosage unit form as used herein refers to a physically discrete unit of therapeutic agent appropriate for the patient to be treated. It will be understood, however, that the total daily usage of the compounds and compositions of the present disclosure will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular patient or organism will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex, and diet of the subject; the time of administration, mute of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts (see, for example, Goodman and Gilman's The Pharmacological Basis of Therapeutics, Tenth Edition, A. Gilman, J. Hardman and L. Limbird, eds., McGraw-Bill Press, 155-173, 2001, which is incorporated herein by reference in its entirety).

Methods of Assessing Responsiveness to Hydroxyurea

[0215] Hydroxyurea, also known as hydroxycarbamide, is an antineoplastic drug for treating proliferative disorders. It is also used in treating sickle cell disease (SCD). Since a SCD patient's responsiveness to hydroxyurea depends on the expression level of HbF induced by hydroxyurea, the responsiveness of RBC can be used to predict one's responsiveness to hydroxyurea.

[0216] To assess whether a patient is responsive to hydroxyurea, a biological sample (e.g., a blood sample) containing RBCs can be collected from the patient and the expression level of a hemoglobin gene, such as a HbF gene, in the RBCs can be measured using a routine method (e.g., real-time PCR). If the expression level of the hemoglobin gene is elevated after treatment of hydroxyurea, this indicates that the patient is responsive to the treatment. On the other hand, if the expression level of the hemoglobin gene is unchanged or decreased, this indicates that the patient might not be responsive to hydroxyurea treatment. The RBCs can be treated by hydroxyurea in vitro. Alternatively, the blood sample can be obtained from a patient who is subject to hydroxyurea treatment.

[0217] A subject identified by the method described herein as inresponsive to a hydroxyurea treatment can be subjected to a different treatment, such as a non-hydroxyurea conventional treatment or using any of the compounds disclosed herein.

[0218] Without further elaboration, it is believed that one skilled in the art can, based on the above description, utilize the present disclosure to its fullest extent. The following specific embodiments are, therefore, to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. All publications cited herein are incorporated by reference for the purposes or subject matter referenced herein.

Examples

Example 1: Effect of Exemplary Compounds in Inducing HbF

[0219] A dual fluorescence reporter assay system was established to screen for the potential HbF-inducing agents. Taking advantage of fluoresence signal detected by fluorometer, these chemical compounds could be surveyed quickly to determine those that can switch-on the fetal globin promoter. Application of this dual fluoresence reporter system has successfully led to identification of several heterocyclic compounds with common core structure (benzo[de]benzo[4,5]imidazo[2,1-a]isoquinolin-7-one) and with higher efficacies/specificities in the induction of the embryonic/fetal globin chains. These chemical compounds may be developed into a new generation of therapeutical drugs for the cure of hemoglobinopathies including sickle cell disease and P-thalassemia (e.g., 3-thalassemia major).

Materials and Methods

[0220] Chemistry. General Information.

[0221] All commercial chemicals and solvents were reagent grade and were used without further purification. Melting points were determined in open capillaries on a Fargo melting point apparatus and are uncorrected. Thin-layer chromatography was performed on silica gel G60 F254 (Merck) with short-wavelength UV light for visualization. High resolution mass spectra were recorded on a Waters HDMS G1 instrument with ESI+, centroid mode, the samples were dissolved in MeOH. High-performance liquid chromatography was performed on Hitachi L-1230 instrument: column: Kinetex 2.6 HILIC (1504.6 mm). Compounds were detected by UV at 260 nm. The mobile phase was MeCN/THF (80:20 v/v) with flow rate of 1 mL/min. The purity of all tested compounds was 98% based on analytical HPLC. 1H NMR spectra was recorded on Bruker AVANCE 600 DRX and/or 400 MHz, Bruker Top-Spin spectrometers in the solvents indicated. The Proton chemical shifts were reported in parts per million ( ppm) relative to (CH.sub.3).sub.4Si and coupling constants (J) in Hertz (Hz) and s, d, t, m, br s, refer to singlet, doublet, triplet, multiplet, broad respectively.

[0222] Chemical Synthesis.

[0223] The synthetic route of preparing exemplified compounds is shown in Schemes 1 and 2. A mixture of naphthalic anhydride and various amines in a molar ratio of 1:10 [such as aniline, dimethylaminopropylamine, dimethylethylene diamine or ethyl 4-aminobutyrate)] in toluene in the presence of acetic acid was heated at reflux. After completion of the reaction, the solvent was evaporated under reduced pressure to dryness. The solid residue was dissolved in ethyl acetate, washed with water, and dried over Na.sub.2SO.sub.4. The solvent was removed by evaporation in vacuo. The amino substituted derivatives were synthesized from the corresponding nitro substituted derivatives by catalytic hydrogenation (5% Pd/C, H.sub.2) at 35 psi. The product was purified by recrystallization with EtOH or by column chromatography.

Compounds

N-Phenyl-1,8-naphthalimide (A1)

[0224] Compound A1 was obtained from 1,8-naphthalic anhydride (1.2 g, 6 mmol) in aniline (50 ml) through general procedure to yield 1.16 g (70.7%), mp 218-220 C. (Lit.sup.16 202 C.) (Jaubert et al., Berichte der deutschen chemischen Gesellschaft. 1895; 28(1):360-364), ESI-HRMS calcd for C.sub.18H.sub.11NO.sub.2 m/z 296.0687 (M+Na), found 296.0692 (M+Na). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.57 (t, J=7.16 Hz, 4H, naphthylic-H) 7.97 (t, J=7.69 Hz, 2H, naphthylic-H) 7.60 (t, J=7.13 Hz, 2H, benzene-H) 7.53 (t, J=7.66 Hz, 1H, benzene-H) 7.45 (d, J=7.27 Hz, 2H, benzene-H).

N-Phenyl-4-nitro-1,8-naphthalimide (A2)

[0225] Compound A2 was obtained from 4-nitro-1,8-naphthalic anhydride (2.43 g, 10 mmol) in toluene (50 ml) and aniline (9.1 ml, 100 mmol) through general procedure to yield 2.775 g (92.7%), mp >280 C. (Lit.sup.17 280-282 C.) (Mitsuo et al., Journal of Synthetic Organic Chemistry, Japan. 1956; 14(9):558-564), FAB-HRMS calcd for C.sub.18H.sub.10N.sub.2O.sub.4 m/z 319.0719 (M+H), found 319.0715 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.83 (d, J=8.55 Hz, 1H, naphthylic-H) 8.71 (d, J=7.27 Hz, 1H, naphthylic-H) 8.69 (d, J=8.04 Hz, 1H, naphthylic-H) 8.66 (d, J=7.42 Hz, 1H, naphthylic-H) 8.2 (t, J=7.43 Hz, 1H, naphthylic-H) 7.61 (t, J=7.23 Hz, 2H, benzene-H) 7.55 (t, J=6.89 Hz, 1H, benzene-H) 7.48 (d, J=7.27 Hz, 2H, benzene-H).

N-(3,4-Dichloro-phenyl)-4-nitro-1,8-naphthalimide (A3)

[0226] Compound A3 was obtained from of 4-nitro-1,8-naphthalic anhydride (2.43 g, 10 mmol) and 3,4-dichloroaniline (16 g, 100 mmol) through general procedure to yield 3.3 g (85.2%), mp >280 C., FAB-HRMS calcd for C.sub.18H.sub.8Cl.sub.2N.sub.2O.sub.4 m/z 386.9939 (M+H), found 386.993 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.79 (d, J=8.33 Hz, 1H, naphthylic-H) 8.67 (d, J=6.62 Hz, 1H, naphthylic-H) 8.64 (d, J=8.12 Hz, 1H, naphthylic-H) 8.6 (d, J=7.91 Hz, 1H, naphthylic-H) 8.15 (t, J=8.65 Hz, 1H, naphthylic-H) 7.86 (d, J=8.55 Hz, 1H, benzene-H) 7.83 (s, 1H, benzene-H) 7.5 (d, J=8.65 Hz, 1H, benzene-H).

N-(3-Methoxy-phenyl)-4-nitro-1,8-naphthalimide (A4)

[0227] Compound A4 was obtained from 4-nitro-1,8-naphthalic anhydride (2.43 g, 10 mmol) and m-anisidine (11.2 ml, 100 mmol) through general procedure to yield 3.21 g (92.2%), mp >280 C., FAB-HRMS calcd for C.sub.19H.sub.12N.sub.2O.sub.5 m/z 349.0824 (M+H), found 349.0832 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.83 (d, J=8.76 Hz, 1H, naphthylic-H) 8.71 (d, J=7.27 Hz, 1H, naphthylic-H) 8.69 (d, J=7.48 Hz, 1H, naphthylic-H) 8.65 (d, J=7.48 Hz, 1H, naphthylic-H) 8.2 (t, J=8.16 Hz, 1H, naphthylic-H) 7.51 (t, J=8.01 Hz, 1H, benzene-H) 7.12 (m, 2H, benzene-H) 7.05 (d, J=8.15 Hz, 1H, benzene-H) 3.85 (s, 3H, OCH.sub.3).

N-(4-Butyric acid-phenyl)-4-nitro-1,8-naphthalimide (A5)

[0228] Compound A5 was obtained from 4-nitro-1,8-naphthalic anhydride (486.4 g, 2 mmol) and 4-(4-amino-phenyl)butyric acid (358.5 mg, 20 mmol) through general procedure to yield 877.5 mg (95.1%), mp 254.5-256 C., ESI-HRMS calcd for C.sub.22H.sub.12N.sub.2O.sub.6 m/z 427.0906 (M+Na), found 427.0898 (M+Na). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.82 (d, J=8.41 Hz, 1H, naphthylic-H) 8.7 (d, J=7.27 Hz, 1H, naphthylic-H) 8.68 (d, J=8.14 Hz, 1H, naphthylic-H) 8.64 (d, J=7.76 Hz, 1H, naphthylic-H) 8.19 (t, J=8.03 Hz, 1H, naphthylic-H) 7.42 (d, J=8.03 Hz, 2H, benzene-H) 7.38 (d, J=8.41 Hz, 2H, benzene-H) 2.75 (t, J=7.84 Hz, 2H, CH.sub.2CH.sub.2CH.sub.2COOH) 2.35 (t, J=7.46 Hz, 2H, CH.sub.2CH.sub.2CH.sub.2COOH) 1.94 (quin, J=7.55 Hz, 2H, CH.sub.2CH.sub.2CH.sub.2COOH).

N-Phenyl-4-chloro-1,8-naphthalimide (A6)

[0229] Compound A6 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and aniline (1.8 ml, 20 mmol) through general procedure to yield 137.9 mg (22.4%), mp 263-265.5 C. (Lit.sup.18 231-233 C.) (Takaaki et al., Fungicidal Compositions; 1973), FAB-HRMS calcd for C.sub.18H.sub.10ClNO.sub.2 m/z 308.0478 (M+H), found 308.048 (M+H). 1H NMR (500 MHz, DMSO-d.sub.6) 8.75 (d, J=7.7 Hz, 1H, naphthylic-H) 8.67 (d, J=6.6 Hz, 1H, naphthylic-H) 8.52 (d, J=7.9 Hz, 1H, naphthylic-H) 8.16 (d, J=7.9 Hz, 1H, naphthylic-H) 8.12 (t, J=7.5 Hz, 1H, naphthylic-H) 7.60 (t, J=7.18 Hz, 2H, benzene-H) 7.53 (t, J=6.89 Hz, 2H, benzene-H) 7.46 (d, J=7.46 Hz, 2H, benzene-H).

N-(3-Methoxy-phenyl)-4-chloro-1,8-naphthalimide (A7)

[0230] Compound A7 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and m-anisidine (2.3 ml, 20 mmol) through general procedure to yield 250 mg (37%), mp >280 C., FAB-HRMS calcd for C.sub.19H.sub.12ClNO.sub.3 m/z 338.0584 (M+H), found 338.0576 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.67 (d, J=8.72 Hz, 1H, naphthylic-H) 8.6 (d, J=6.47 Hz, 1H, naphthylic-H) 8.45 (d, J=7.9 Hz, 1H, naphthylic-H) 8.08 (d, J=7.9 Hz, 1H, naphthylic-H) 8.05 (t, J=8.71 Hz, 1H, naphthylic-H) 7.43 (t, J=8.01 Hz, 1H, benzene-H) 7.04 (d, J=8.09 Hz, 1H, benzene-H) 7.02 (s, J=7.46 Hz, 1H, benzene-H) 6.96 (d, J=8.72 Hz, 1H, benzene-H) 3.78 (s, 3H, OCH.sub.3).

N-(3-Trifluoromethyl-phenyl)-4-chloro-1,8-naphthalimide (A8)

[0231] Compound A8 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and 3-aminobenzotrifluoride (2.5 ml, 20 mmol) through general procedure to yield 130.2 mg (17.3%), mp 278-279 C., FAB-HRMS calcd for C.sub.19H.sub.9ClF.sub.3NO.sub.2 m/z 376.0352 (M+H), found 376.0345 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.7 (d, J=8.5 Hz, 1H, naphthylic-H) 8.62 (d, J=6.61 Hz, 1H, naphthylic-H) 8.47 (d, J=7.9 Hz, 1H, naphthylic-H) 8.11 (d, J=7.9 Hz, 1H, naphthylic-H) 8.07 (t, J=7.8 Hz, 1H, naphthylic-H) 7.9 (s, 1H, benzene-H) 7.87 (d, J=7.7 Hz, 1H, benzene-H) 7.8 (d, J=7.78 Hz, 1H, benzene-H) 7.77 (t, J=5.83 Hz, 1H, benzene-H).

N-(3-Bromo-phenyl)-4-chloro-1,8-naphthalimide (A9)

[0232] Compound A9 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and 3-bromoaniline (2.2 ml, 20 mmol) through general procedure to yield 102.2 mg (13.2%), mp >280 C., FAB-HRMS calcd for C.sub.18H.sub.9BrClNO.sub.2 m/z 385.9583 (M+H), found 385.9577 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.7 (d, J=8.33 Hz, 1H, naphthylic-H) 8.61 (d, J=7.27 Hz, 1H, naphthylic-H) 8.46 (d, J=7.91 Hz, 1H, naphthylic-H) 8.1 (d, J=7.91 Hz, 1H, naphthylic-H) 8.07 (t, J=7.91 Hz, 1H, naphthylic-H) 7.72 (br s, 1H, benzene-H) 7.7 (d, J=8.12 Hz, 1H, benzene-H) 7.51 (t, J=7.8 Hz, 1H, benzene-H) 7.46 (d, J=7.09 Hz, 1H, benzene-H).

N-(3-Chloro-phenyl)-4-chloro-1,8-naphthalimide (A10)

[0233] Compound A10 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and 3-chloroaniline (2.13 ml, 20 mmol) through general procedure to yield 82.8 mg (12.1%), mp 273-275 C., FAB-HRMS calcd for C.sub.18H.sub.9Cl.sub.2NO.sub.2 m/z 342.0089 (M+H), found 342.0098 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.7 (d, J=8.55 Hz, 1H, naphthylic-H) 8.62 (d, J=7.27 Hz, 1H, naphthylic-H) 8.47 (d, J=7.91 Hz, 1H, naphthylic-H) 8.1 (d, J=7.91 Hz, 1H, naphthylic-H) 8.07 (t, J=7.91 Hz, 1H, naphthylic-H) 7.58 (m, 3H, benzene-H) 7.41 (d, J=5.94 Hz, 1H, benzene-H). Papenfuhs et al., United Kingdom: Hoechst Aktiengesellschaft (Frankfurt am Main, DE); 1977.

N-(3-Bromo-4-fluoro-phenyl)-4-chloro-1,8-naphthalimide (A11)

[0234] Compound A11 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and 3-bromo-4-fluroaniline (3.8 g, 20 mmol) through general procedure to yield 398 mg (49.2%), mp >280 C., FAB-HRMS calcd for C.sub.18H.sub.8BrClFNO.sub.2 m/z 403.9489 (M+H), found 403.949 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.69 (d, J=8.76 Hz, 1H, naphthylic-H) 8.61 (d, J=7.29 Hz, 1H, naphthylic-H) 8.46 (d, J=7.91 Hz, 1H, naphthylic-H) 8.1 (d, J=7.91 Hz, 1H, naphthylic-H) 8.07 (t, J=7.65 Hz, 1H, naphthylic-H) 7.87 (dd, J=6.3, 2.24 Hz, 1H, benzene-H) 7.54 (m, 2H, benzene-H)

N-(4-Fluoro-phenyl)-4-chloro-1,8-naphthalimide (A12)

[0235] Compound A12 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and 4-fluroaniline (1.9 ml, 20 mmol) through general procedure to yield 264 mg (40.5%), mp 274-276 C., FAB-HRMS calcd for C.sub.18H.sub.9ClFNO.sub.2 m/z 326.0384 (M+H), found 326.0391 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.69 (d, J=8.56 Hz, 1H, naphthylic-H) 8.61 (d, J=7.34 Hz, 1H, naphthylic-H) 8.46 (d, J=7.91 Hz, 1H, naphthylic-H) 8.1 (d, J=7.91 Hz, 1H, naphthylic-H) 8.06 (t, J=8.79 Hz, 1H, naphthylic-H) 7.46 (m, 2H, benzene-H) 7.37 (m, 2H, benzene-H)

N-(4-Bromo-phenyl)-4-chloro-1,8-naphthalimide (A13)

[0236] Compound A13 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and 4-bromoaniline (3.4 g, 20 mmol) through general procedure to yield 513 mg (66.3%), mp >280 C., FAB-HRMS calcd for C.sub.18H.sub.9BrClNO.sub.2 m/z 385.9583 (M+H), found 385.9587 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.69 (d, J=7.38 Hz, 1H, naphthylic-H) 8.61 (d, J=7.95 Hz, 1H, naphthylic-H) 8.46 (d, J=7.91 Hz, 1H, naphthylic-H) 8.1 (d, J=7.91 Hz, 1H, naphthylic-H) 8.06 (t, J=8.42 Hz, 1H, naphthylic-H) 7.74 (d, J=8.52 Hz, 2H, benzene-H) 7.39 (d, J=8.51 Hz, 2H, benzene-H).sup.19

N-(3,4-Dichloro-phenyl)-4-chloro-1,8-naphthalimide (A14)

[0237] Compound A14 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and 3,4-dichloroaniline (3.24 g, 20 mmol) through general procedure to yield 123 mg (15.9%), mp >280 C. (Lit.sup.18 >300 C.), FAB-HRMS calcd for C.sub.18H.sub.8Cl.sub.3NO.sub.2 m/z 375.9699 (M+H), found 375.9697 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.7 (d, J=8.33 Hz, 1H, naphthylic-H) 8.62 (d, J=6.62 Hz, 1H, naphthylic-H) 8.47 (d, J=7.91 Hz, 1H, naphthylic-H) 8.11 (d, J=7.91 Hz, 1H, naphthylic-H) 8.07 (t, J=7.91 Hz, 1H, naphthylic-H) 7.85 (s, 1H, benzene-H) 7.82 (m, 1H, benzene-H) 7.48 (d, J=8.12 Hz, 1H, benzene-H).

N-(2-Dimethylamino-ethyl)-1,8-naphthalimide hydrochloride (B1)

[0238] Compound B1 was obtained from 1,8-naphthalic anhydride (1 g, 5.05 mmol) and N,N-dimethylethylene diamine (5.5 ml, 50 mmol) through general procedure and form water soluble salt with 1.75 M HCl/EtOAc (2.9 ml, 5.1 mmol) to yield 1.3 g (84.5%), mp >280 C. (Lit.sup.20 296-298 C.), ESI-HRMS calcd for C.sub.16H.sub.17ClN.sub.2O.sub.2 m/z 303.09 (MH), found 303.0906 (MH). .sup.1H NMR (500 MHz, Methanol-d4) 9.61 (br s, 1H, HCl) 8.53 (d, J=7.13, 2H, naphthylic-H) 8.51 (d, J=7.47 Hz, 2H, naphthylic-H) 7.91 (t, J=7.47 Hz, 2H, naphthylic-H) 4.41 (t, J=5.98 Hz, 2H, ethylene-CH.sub.2) 3.45 (t, J=5.98 Hz, 2H, ethylene-CH.sub.2) 3.03 (s, 6H, N(CH.sub.3).sub.2).

N-(2-Dimethylamino-ethyl)-4-nitro-1,8-naphthalimide (B2)

[0239] Compound B2 was obtained from 4-nitro-1,8-naphthalic anhydride (1 g, 4.11 mmol) and N,N-dimethylethylene diamine (4.37 ml, 40 mmol) through general procedure to yield 843.6 mg (65.7%), mp 131-133 C. (Lit.sup.20 106-109 C.), ESI-HRMS calcd for C.sub.16H.sub.15N.sub.3O.sub.4 m/z 314.1141 (M+H), found 314.1148 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.72 (d, J=8.55 Hz, 1H, naphthylic-H) 8.64 (d, J=7.27 Hz, 1H, naphthylic-H) 8.62 (d, J=8.12 Hz, 1H, naphthylic-H) 8.56 (d, J=8.12 Hz, 1H, naphthylic-H) 8.1 (t, J=8.01 Hz, 1H, naphthylic-H) 4.16 (t, J=6.95 Hz, 2H, ethylene-CH.sub.2) 2.53 (m, 2H, ethylene-CH.sub.2) 2.38 (s, 6H, N(CH.sub.3).sub.2).

N-(3-Dimethylamino-propyl)-4-nitro-1,8-naphthalimide (B3)

[0240] Compound B3 was obtained from 4-nitro-1,8-naphthalic anhydride (1 g, 4.11 mmol) and 3-dimethylaminopropylamine (4.4 ml, 40 mmol) through general procedure to yield 564 mg (42.0%), mp 108-110 C. (Lit.sup.21 106-109 C.), ESI-HRMS calcd for C.sub.17H.sub.17N.sub.3O.sub.4 m/z 328.1297 (M+H), found 328.1299 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.71 (d, J=8.55 Hz, 1H, naphthylic-H) 8.64 (d, J=7.27 Hz, 1H, naphthylic-H) 8.61 (d, J=7.91 Hz, 1H, naphthylic-H) 8.55 (d, J=7.91 Hz, 1H, naphthylic-H) 8.1 (t, J=7.91 Hz, 1H, naphthylic-H) 4.08 (t, J=7.25 Hz, 2H, propylene-CH.sub.2) 2.31 (t, J=6.95 Hz, 2H, propylene-CH.sub.2) 2.12 (s, 6H, N(CH.sub.3).sub.2) 1.78 (quin, J=7.27 Hz, 2H, propylene-CH.sub.2).

N-(2-dimethylamino-ethyl)-4-chloro-1,8-naphthalimide hydrochloride (B4)

[0241] Compound B4 was obtained from 4-chloro-1,8-naphthalic anhydride (1 g, 4.3 mmol) and N,N-dimethylethylene diamine (4.37 ml, 40 mmol) through general procedure and form water soluble salt with 1.75 M HCl/EtOAc (2.6 ml, 4.5 mmol) to yield 1.16 g (81.5%), mp >280 C. (Lit.sup.22 293-295 C.), ESI-HRMS calcd for C.sub.16H.sub.16Cl.sub.2N.sub.2O.sub.2 m/z 332.1166 (MH), found 332.1157 (MH). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 9.76 (br s, 1H, HCl) 8.66 (d, J=8.55 Hz, 1H, naphthylic-H) 8.62 (d, J=7.27 Hz, 1H, naphthylic-H) 8.47 (d, J=7.91 Hz, 1H, naphthylic-H) 8.09 (d, J=7.91 Hz, 1H, naphthylic-H) 8.05 (t, J=7.52 Hz, 1H, naphthylic-H) 4.4 (t, J=5.98 Hz, 2H, ethylene-CH.sub.2) 3.44 (t, J=5.88 Hz, 2H, ethylene-CH.sub.2) 2.88 (s, 6H, N(CH.sub.3).sub.2).

N-Ethyl-1,8-naphthalimide (C1)

[0242] Compound C1 was obtained from 1,8-naphthalic anhydride (396.4 mg, 2 mmol) and ethylamine (12.9, 20 mmol) through general procedure to yield 168 mg (37.3%), mp 171-172.5 C. (Lit.sup.18 158 C.), FAB-HRMS calcd for C.sub.14H.sub.11NO.sub.2 m/z 226.0868 (M+H), found 226.0868 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.57 (d, J=7.99 Hz, 2H, naphthylic-H) 8.53 (d, J=8.67 Hz, 2H, naphthylic-H) 7.94 (t, J=7.69 Hz, 2H, naphthylic-H) 4.16 (q, J=7.05 Hz, 2H, CH.sub.2CH.sub.3) 1.28 (t, J=7.05 Hz, 3H, CH.sub.2CH.sub.3).

N-Methyl-4-nitro-1,8-naphthalimide (C2)

[0243] Compound C2 was obtained from 4-nitro-1,8-naphthalic anhydride (2.43 g, 10 mmol) and methylamine (77 g, 100 mmol) through general procedure to yield 1.31 g (51.1%), mp 209-212.5 C. (Lit.sup.18 208-209 C.), FAB-HRMS calcd for C.sub.13H.sub.8N.sub.2O.sub.4 m/z 257.0562 (M+H), found 257.0563 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.7 (d, J=8.22 Hz, 1H, naphthylic-H) 8.62 (m, 2H, naphthylic-H) 8.56 (m, 1H, naphthylic-H) 8.09 (t, J=8.22 Hz, 1H, naphthylic-H) 3.41 (s, 3H, CH.sub.3).

N-Ethyl-4-nitro-1,8-naphthalimide (C3)

[0244] Compound C3 was obtained from 4-nitro-1,8-naphthalic anhydride (2.43 g, 10 mmol) and ethylamine (64.4 g, 40 mmol) through general procedure to yield 922.6 mg (34.1%), mp 191-192.5 C. (Lit.sup.18 187.5-188.5 C.), FAB-HRMS calcd for C.sub.14H.sub.10N.sub.2O.sub.4 m/z 271.0719 (M+H), found 271.0722 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.71 (d, J=9.0 Hz, 1H, naphthylic-H) 8.63 (m, 2H, naphthylic-H) 8.56 (m, 1H, naphthylic-H) 8.09 (t, J=7.63 Hz, 1H, naphthylic-H) 4.09 (q, J=7.04 Hz, 2H, CH.sub.2CH.sub.3) 1.24 (t, J=7.04 Hz, 3H, CH.sub.2CH.sub.3).

N-Methyl-4-chloro-1,8-naphthalimide (C4)

[0245] Compound C4 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and methylamine (15.5 g, 200 mmol) through general procedure to yield 160 mg (32.6%), mp 186-188 C. (Lit.sup.18 171-173 C.) (Takaaki et al., Fungicidal Compositions; 1973), FAB-HRMS calcd for C.sub.13H.sub.8ClNO.sub.2 m/z 246.032 (M+H), found 246.0318 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.62 (d, J=8.69 Hz, 1H, naphthylic-H) 8.60 (d, J=7.48 Hz, 1H, naphthylic-H) 8.45 (d, J=7.69 Hz, 1H, naphthylic-H) 8.06 (d, J=7.91 Hz, 1H, naphthylic-H) 8.02 (t, J=7.91 Hz, 1H, naphthylic-H) 3.4 (s, 3H, CH.sub.3).

N-Ethyl-4-chloro-1,8-naphthalimide (C5)

[0246] Compound C5 was obtained from 4-chloro-1,8-naphthalic anhydride (465 mg, 2 mmol) and ethylamine (12.9 g, 200 mmol) through general procedure to yield 170 mg (32.7%), mp 167-170 C. (Lit.sup.18 165-166 C.) (Takaaki et al., Fungicidal Compositions; 1973), FAB-HRMS calcd for C.sub.14H.sub.10ClNO.sub.2 m/z 260.0478 (M+H), found 260.0484 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.61 (t, J=7.91 Hz, 2H, naphthylic-H) 8.45 (d, J=7.91 Hz, 1H, naphthylic-H) 8.05 (d, J=7.91 Hz, 1H, naphthylic-H) 8.02 (t, J=7.49 Hz, 1H, naphthylic-H) 4.08 (q, J=7.05 Hz, 2H, CH.sub.2CH.sub.3) 1.22 (t, J=7.05 Hz, 3H, CH.sub.2CH.sub.3).

N-(Butyric acid ethyl ester)-4-nitro-1,8-naphthalimide (D1)

[0247] Compound D1 was obtained from ethyl 4-aminobutyrate hydrochloride (8.4 g, 50 mmol) and 4-nitro-1,8-naphthalic anhydride (1.22 g, 5 mmol) through general procedure to yield 1.65 g (97.0%), mp 115-116 C., FAB-HRMS calcd for C.sub.18H.sub.16N.sub.2O.sub.6 m/z 357.1087 (M+H), found 357.1091 (M+H). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.72 (d, J=8.55 Hz, 1H, naphthylic-H) 8.64 (d, J=7.27 Hz, 1H, naphthylic-H) 8.62 (d, J=8.12 Hz, 1H, naphthylic-H) 8.56 (d, J=7.91 Hz, 1H, naphthylic-H) 8.1 (t, J=8.01 Hz, 1H, naphthylic-H) 4.11 (t, J=6.84 Hz, 2H, CH.sub.2CH.sub.2CH.sub.2COOCH.sub.2CH.sub.3) 3.97 (q, J=7.05 Hz, 2H, CH.sub.2CH.sub.2CH.sub.2COOCH.sub.2CH.sub.3) 2.41 (t, J=7.37 Hz, 2H, CH.sub.2CH.sub.2CH.sub.2COOCH.sub.2CH.sub.3) 1.94 (quin, J=7.11 Hz, 2H, CH.sub.2CH.sub.2CH.sub.2COOCH.sub.2CH.sub.3) 1.12 (t, J=7.05 Hz, 3H, CH.sub.2CH.sub.2CH.sub.2COOCH.sub.2CH.sub.3)

N-Phenyl-4-amino-1,8-naphthalimide (E1)

[0248] Compound E1 was obtained from N-phenyl-4-nitro-naphthalimide (1.27 g, 4 mmol) through general procedure to yield 583 mg (56.4%), mp 276-278 C. (Lit.sup.23 302-304 C.) (Mitsuo et al., Journal of Synthetic Organic Chemistry, Japan. 1956; 14(8):504-508), ESI-HRMS calcd for C.sub.18H.sub.12N.sub.2O.sub.2 m/z 287.0821 (MH), found 287.0816 (MH). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 8.66 (d, J=7.69 Hz, 1H, naphthylic-H) 8.43 (d, J=6.62 Hz, 1H, naphthylic-H) 8.19 (d, J=8.33 Hz, 1H, naphthylic-H) 7.69 (t, J=8.58 Hz, 1H, naphthylic-H) 7.5 (m, 4H, benzene-H, amino-H) 7.43 (t, J=7.8 Hz, 1H, benzene-H) 7.3 (d, J=6.95 Hz, 2H, benzene-H) 6.88 (d, J=8.33 Hz, 1H, naphthylic-H).

N-(2-dimethylamino-ethyl)-4-amino-1,8-naphthalimide hydrochloride (E2)

[0249] Compound E2 was obtained from N-(2-dimethylamino-ethyl)-4-nitro-1,8-naphthalimide (200 mg, 0.64 mmol) through general procedure and form water soluble salt with 1.75 M HCl/EtOAc (0.37 ml, 0.65 mmol) to yield 174.2 mg (77.3%), mp >280 C. (Lit.sup.20 184-185 C.) (Norton et al., Anticancer Drugs. 2008; 19(1):23-36), ESI-HRMS calcd for C.sub.16H.sub.18ClN.sub.3O.sub.2 m/z 318.1009 (MH), found 318.1003 (MH). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 9.3 (br s, 1H, HCl) 8.66 (d, J=8.72 Hz, 1H, naphthylic-H) 8.46 (d, J=6.79 Hz, 1H, naphthylic-H) 8.22 (d, J=8.55 Hz, 1H, naphthylic-H) 7.68 (t, J=8.73 Hz, 1H, naphthylic-H) 7.56 (br s, 2H, exchangeable, amino-H) 6.87 (d, J=8.33 Hz, 1H, naphthylic-H) 4.35 (t, J=5.88 Hz, 2H, ethylene-CH.sub.2) 3.42 (in H.sub.2O, show in methanol-d4, ethylene-CH.sub.2) 2.89 (br s, 6H, N(CH.sub.3).sub.2). Norton et al., Anticancer Drugs. 2008; 19(1):23-36.

N-(3-dimethylamino-propyl)-4-amino-1,8-naphthalimide hydrochloride (E3)

[0250] Compound E3 was obtained from N-(3-dimethylamino-propyl)-4-nitro-naphthalimide (250 mg, 0.76 mmol) through general procedure and form water soluble salt with 1.75 M HCl/EtOAc (0.45 ml, 0.8 mmol), to yield 120 mg (42.8%), mp >280 C. (Lit.sup.21 184-185 C.) (Stevenson et al., J Med Chem. 1984; 27(12):1677-1682), ESI-HRMS calcd for C.sub.17H.sub.20ClN.sub.3O.sub.2 m/z 303.09 (M-Cl), found 303.0903 (M-Cl). .sup.1H NMR (500 MHz, DMSO-d.sub.6) 9.75 (br s, 1H, HCl) 8.65 (d, J=8.79 Hz, 1H, naphthylic-H) 8.44 (d, J=8.79 Hz, 1H, naphthylic-H) 8.21 (d, J=8.33 Hz, 1H, naphthylic-H) 7.67 (t, J=8.79 Hz, 1H, naphthylic-H) 7.51 (br s, 2H, exchangeable, amino-H) 6.86 (d, J=8.55 Hz, 1H, naphthylic-H) 4.08 (t, J=6.62 Hz, 2H, propylene-CH.sub.2) 3.11 (t, J=7.82 Hz, 2H, propylene-CH.sub.2) 2.73 (s, 6H, N(CH.sub.3).sub.2) 2.02 (m, 2H, propylene-CH.sub.2).

[0251] The Primary Erythroid Cell Culture.

[0252] The peripheral blood samples were purchased from the Taipei Blood Center. The concentrated blood was diluted 1:5 (V/V) with PBS and gentle layer on Ficoll-Hypaque (d=1.007 g/ml) (Ge-Healthcare). After centrifugation at 400 g for 20 minutes, the cells in the inter-phase region were collected. The collected cells were washed with PBS and centrifuged at low speed for three times. The remained mononuclear cells were expanded in Phase I medium containing 1SFEM (StemSpan), 100 ng/ml SCF, 20 ng/ml IL-3, 20 ng/ml IL-6, and 100 ng/ml Fit3-L at 37 C. incubator with 5% CO.sub.2 for 7 days. The expanded mononuclear cells were then differentiated in Phase II medium which contains 1SFEM (StemSpan), 20 ng/ml SCF, 5 ng/ml IL-3, 1 U/ml EPO for another 7 days. The resulted differentiated erythroid cells were treated with indicated compounds of different dosages with the seeding density of 510.sup.5 cells/ml for another 3 days.

[0253] Quantitative RT-PCR.

[0254] After 3 days of drug-treatment, the total RNA was extracted by RNA Spin mini kit (Ge-Healthcare) and reverse-transcription was performed by using superscript II (Invitrogen) followed the manufactors instruction. Quantitative PCR was performed on LightCycler with using STBR green master mix followed the manufactors instruction (Roche). The absolute quantitative RT-PCR was calculated by the standard curve with supplying a known copy number of plasmids. The relative quantitative RT-PCR was normalized to the expression levels of the -actin and compared to the mock control.

[0255] Cell Viability Assay.

[0256] The cell viability was detected by using AlamarBlue reagent (Invitrogen). After 3 days, the drug-tested cells were transferred into 96-well plate, added with 1/10 volume of AlamarBlue reagent, and then incubate at 37 C. overnight. The cell viability was calculated by a fluorescence reader (Ex 530-560 nm, Em 590 nm).

[0257] Western Blot Analysis.

[0258] After 3 days of compound treatment, total protein or histone of the drug-treated cells was extracted. Histone was extracted by histone extraction buffer (10 mM HEPES, 1.5 mM MgCl.sub.2, 10 mM KCl, 0.5 mM DTT, 1.5 mM PMSF and 0.2 N HCl) at 4 C. overnight then centrifuged at 13,000 rpm for 5 minutes and collected the supernatant. Thirty microgram of histone extracts were separated in 15% SDS-PAGE and blotted onto PVDF membrane. After blocking with 5% non-fat milk in TBST, the membrane was then incubated with primary antibody against total histone H4 or total acetyl histone H4 at 4 C. overnight. Total protein was extracted by modified RIPA (50 mM Tris-HCl pH7.8, 150 mM NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, 1 mM EGTA, 5 mM EDTA, 10 mM NaF, 1 mM NaV.sub.3O.sub.4 and 1 complete, EDTA-free Protease Inhibitor Cocktail). Thirty microgram of total protein extracts were separate by 10% SDS-PAGE and blotted onto PVDF membrane. After blocking with 5% non-fat milk in TBST, the membrane was incubated with primary antibody against BCL11A, p38, p-p38, -globin or -actin at 4 C. overnight. After incubating with horseradish peroxidase-conjugated secondary antibodies and washing the blot, the signals of indicated proteins were visualized by using ECL (PerKinElmer) following the manufactor's protocol.

[0259] Statistical Analysis.

[0260] The data are presented as the mean plus or minus standard error of the mean (SEM) for at least 3 experiments. Statistical analysis of the raw data was performed by the 2-tailed t test. The Student t test was used to measure differences in samples. A probability of less than 0.05 (P<0.05) was considered significant. (*: P<0.05, **: P<0.01, ***: P<0.001)

Results

N-Substituted 1,8-Naphthalimide Derivatives are Potential HbF-Inducing Agents.

[0261] To search for new agents, which are capable of inducing the expression of globin, a series of naphthalimide derivatives with various substituent(s) on the naphthalene ring and the heterocyclic N-atom (A1-A14, B1-B4, C1-C5, D1, and E1-E3, FIG. 2) were synthesized. These derivatives were subjected to evaluating the globin induction abilities in differentiated peripheral blood mononuclear cells (PBMCs). The PBMCs isolated from the peripheral blood of donors were expanded and differentiated for a total of 14 days in a two-phase liquid culture. The resulted primary erythoid cells were then treated with different dosages of naphthalimide derivatives individually for another 3 days. The expression levels of the endogenous globin gene were estimated by relative quantitative RT-PCR and the proliferation rates of these cells were examined by AlamarBlue cell viability assay. Among these naphthalimide derivatives, compounds A2, B1, B4, E1, E2, and E3 were found to have potent globin-inducing effect; 2-4.5-folds induction of the globin gene expression in the compounds-treated cells (FIG. 3).

[0262] The proliferation rates of these cells displayed reverse relationships with the concentrations of these compounds. To compare the HbF-inducing capability of these compounds, the induction folds of globin gene at concentrations of their IC.sub.50 (the half maximal inhibitory concentration) were examined. The IC.sub.50 values indicate that the concentrations of the individual compound at which the cell (primary human erythroid cells) proliferation rate is reduced by 50%. As shown in Table 3, compounds B1, B4, E2, and E3 significantly elevated the expression levels of globin mRNA up to 4.1-, 3.3-, 4.0-, and 2.7-folds at concentrations of their IC.sub.50, respectively (Table 3), While, compounds A2 and E1 displayed only minor globin-inducing potentials with only 1.1- and 1.3-folds of the globin gene induction at concentrations of their IC.sub.50, respectively.

TABLE-US-00004 TABLE 3 Comparison of IC.sub.50, solubility, and globin gene inducing abilities of globin-inducing compounds. EC (M) induction globin induction Compound ID DMSO H.sub.2O IC.sub.50(M) of HU IC.sub.50(2.3 fold) IC.sub.50/EC (IC.sub.50) S51021 10 mM 21.8 2.3 9.5 2.97 A1 10 mM 2.3 7.8 0.3 1.3 B1 >100 mM 10.7 4.7 2.3 4.1 B4 >100 mM 5.0 2.2 2.3 3.3 E1 10 mM 0.3 2.9 0.1 1.1 E2 (SS-2394) >100 mM 2.3 0.8 2.9 4.0 E3 >100 mM 11.2 7.4 1.5 2.7 HU >100 mM 145.9 145.9 1 2.3 NaB >500 mM 221.6 ND ND 1.3

[0263] The primary human erythorid cells were treated with parental compound, NaB, HU, or synthesized naphthalimide derivatives (A1, B1, B4, E, E2, or E3) individually. The values of IC.sub.50 (the half mximal inhibitory concentration), EC (effective concentration), and the ratio of IC.sub.50 to EC (IC.sub.50/EC) were calculated, which were used to compare the advantages of these g globin-inducing compounds to hydroxyurea and each other. The g globin induction fold of each compound at its IC.sub.50 was also shown. ND: non-detectable.

Comparisons of the Therapeutic Potentials Among Naphthalimide Derivatives, S51021, NaB, and HU

[0264] In order to evaluate the therapeutic potentials of naphthalimide derivatives A2, B1, B4, E1, E2, and E3 in comparison with the parental S51021 as well as other HbF inducers (e.g., NaB and HU), the effective concentration (EC) of each compound was investigated first. The EC values are defined as the concentrations of compounds that induce the globin gene expression by 2.3-folds, the fold of the globin gene induction by HU at the concentration of IC.sub.50. The ratio of IC.sub.50 to EC (IC.sub.50/EC) is used to determine whether the compound tested has superior therapeutic effect than that of HU (the ratio IC.sub.50/EC of HU was set as 1). Although compounds A2 and E1 can activate the globin gene expression, their therapeutic potentials were lower than that of HU. In contrast, compounds B1, B4, E2 and E3 exhibit better therapeutic potentials than HU. Of these agents, compound E2 (SS-2394) has the highest IC.sub.50/EC ratio (IC.sub.50/EC=2.79) and relatively higher globin induction folds (4.0-folds) at the concentration of IC.sub.50 (Table I). Although none of naphthalimide derivatives showed better IC.sub.50/EC value than that of S51021, compounds B1, B4, and E2 indeed displayed higher globin induction folds than that of S51021 at the concentrations of their IC.sub.50. It should be noted that compounds B1, B4, E2, and E3 were synthesized as hydrochloride salts and have greatly improved water-solubility (Table 3).

The HbF-Inducing Agents Specifically Induce the Embryonic/Fetal Globin Genes Expression.

[0265] To evaluate the specificity of S51021, SS-2394, HU, and NaB in mediating globin genes activation, the amounts of individual P3-liked globin mRNAs were measured by absolute quantitative RT-PCR. As shown in Table 4, the proportions of the embryonic E globin and the fetal globin mRNAs were both significantly increased by the tested compounds. On the other hand, the proportions of the globin mRNA were notably reduced after drug-treatment. Among them, the SS-2394 is the most effective fetal globin inducer; increasing the proportion of the fetal globin from 7.9% to 11.6% in the drug-treated primary human adult erythroid cells, whereas the proportion of the adult globin mRNA was reduced from 88.8% to 82.3% (Table 4).

TABLE-US-00005 TABLE 4 The expression levels of each -like globin chains in the primary human erythroid cells globin globin globin globin Mock 0.0 0.0% 7.9 0.8% 88.8 1.0% 3.3 0.2% HU 0.1 0.0% 11.2 1.1% 84.2 1.3% 4.5 0.2% NaB 0.1 0.0% 10.8 1.0% 87.4 1.0% 1.8 0.1% S51021 0.1 0.0% 10.6 1.2% 85.6 1.2% 3.7 0.2% SS-2394 0.5 0.1% 11.6 1.0% 82.3 1.1% 5.5 0.4%

[0266] The primary human erythroid cells were treated with HU (145.9 M), NaB (221.6 M), S51021 (21.8 M) or SS2394 (2.3 M) for 3 days, and the expression levels of each globin mRNA were estimated by absolute quantitative RT-PCR analysis. The proportions of individual 3-like globin chains among the total 3-like globin mRNA are shown. Data are presented as the meanSEM, N=3.

Expression Levels of Transcription Activator and Repressor are Modulated by the HbF-Inducing Compounds

[0267] It has been reported that several transcriptional factors, such as NF-E4, c-Myb, and BCL11A, are involved in the regulation of the globin gene expression. Zhou et al., Mol Cell Biol. 2000; 20(20):7662-7672; Jiang et al., Blood. 2006; 108(3): 1077-1083; and Sankaran et al., Science. 2008; 322(5909):1839-1842. NF-E4 was reported to form transcription activation complex, stage selector protein (SSP), which recruited to the globin promoter and switched on the globin gene expression in the primary human adult erythorid cells. Zhou et al., Mol Cell Biol. 2000; 20(20):7662-7672. c-Myb was showed to involve in erythropoiesis and its overexpression in K562 cells inhibited the -globin gene expression..sup.25 More recently, BCL11A was identified to be a developmental stage-specific repressor for controlling the globin gene expression. Sankaran et al., Science. 2008; 322(5909):1839-1842. The expression level of full-length BCL11A was increased in parallel with globin switching, suggesting that BCL11A represses the globin gene expression in the adult erythroid cells. To clarify the regulatory factors involved in modulating globin gene expression in the drug-treated primary human adult erythroid cells, the mRNA levels of the globin as well as these transcription factors in response to compounds treatment were determined by the relative quantitative RT-PCR (FIG. 4A). The expression levels of the globin mRNA were significantly up-regulated to 1.9-3.7-folds in the cells treated with S51021, SS-2394, or HU, but not NaB (FIG. 4A).

[0268] The elevation of protein expression levels of the globin chain were further confirmed in the primary human adult erythroid cells treated with S51021 and SS-2394 (FIG. 4B). The expression levels of NF-E4 were increased to 2.4- and 3.4-folds in the cells treated with S51021 and SS-2394, respectively, but decreased to 0.6 fold in the cells treated with either HU or NaB (FIG. 4A). The expression level of c-Myb mRNA was reduced only in the cells treated with S51021 (FIG. 4A). The expression levels of BCL11A mRNA were significantly reduced to 0.8-, 0.6-, and 0.6-folds by S51021, SS-2394, and NaB, respectively (FIG. 4A). A significant reduction of BCL11A protein was also revealed in the primary human adult erythroid cells treated with S51021, SS-2394, or NaB (FIG. 4C). Taken together, these data suggest that the modulations of certain transcription factors, such as NF-E4, c-Myb, and BCL11A, may cooperatively contribute to the globin gene induction by the tested compounds. Furthermore, both S51021 and SS-2394 can efficiently induce the re-activation of globin gene in the primary human adult erythroid cells, which were not responded upon HU treatment, indicating that S51021 and SS-2394 activate the globin gene expression through a distinct mechanism from that of HU (FIG. 4D).

p38 Signaling Pathway is Involved in the Globin Gene Activation Triggered by the HbF-Inducing Agents.

[0269] In addition to these transcription factors, the activation of p38 MAPK signaling pathway was also suggested to mediate the fetal globin gene expression in response to NaB or TSA treatment. Ramakrishnan et al., Blood Cells Mol Dis; 47(1): 12-22; and Pace et al., Exp Hematol. 2003; 31(11):1089-1096. To verify whether the p38 MAPK signaling pathway is involved in the activation of the globin expression by the tested compounds, the phosphorylation status of p38 in these drug-treated erythroid cells was examined by Western blot analysis. Our data showed that all the tested compounds (S51021, SS-2394, HU, and NaB) significantly increased the phosphorylation levels of p38 in parallel with the elevated globin protein (FIG. 5), suggesting that the activation of p38 MAPK signaling pathway may, at least in part, contribute to the re-activation of the globin gene expression by these HbF-inducing compounds.

S51021 and SS-2394 are not Histone Deacetylase Inhibitors.

[0270] Several HDACi, such as NaB, apicidin, and TSA, have been reported to function as HbF-inducing agents. General believes that the hyper-acetylation of histone by HDACi treatments would contribute to the activation of the globin gene expression. To realize whether the epigenetic modification of histone H4 is manipulated by S51021 or SS-2394 treatment, primary human adult erythroid cells were treated with S51021, SS-2394, HU, or NaB for 3 days, and the acetylation status of cellular histone H4 was then analyzed (FIG. 6). Our data showed that the global acetylation status of histone H4 (total acH4) was significantly enhanced in the NaB-treated primary erythroid cells. In the contrast, the amounts of global acetylated histone H4 were not dramatically alternated by S51021, SS-2394 and HU. It indicates that the S51021, SS-2394, and HU are not histone deacetylase inhibitors.

DISCUSSION

Underlying Mechanisms Involved in Reactivation of the Globin Gene Expression Mediated by S51021 and SS-2394

[0271] Unlike NaB or TSA, S51021 and SS-2394 are not histone deacetylase inhibitors, which did not modulate the global acetylation statues of histone H4 (FIG. 6). Furthermore, it was found that both S51021 and SS-2394 could efficiently re-activate the expression of globin gene even in these primary erythroid cells which were not responded to HU (FIG. 4D). All these data demonstrate that the S51021 and SS-2394 belong to a novel class of HbF-inducing agents, which are functional different from these of previous identified compounds including NaB, HU, and TSA. Although the detail mechanisms involved in the globin gene induction have not yet been completely clarified, our current studies provide insight into understanding of the underlying mechanism that involves in the re-activation of the globin gene expression mediated by S51021 or SS-2394. In the primary human adult erythroid cells, S51021 and SS-2394 treatment significantly down-regulated the expression levels of transcription repressor BCL11A. On the other hand, the expression level of transcription activator NF-E4 was significantly up-regulated by S51021 and SS-2394 treatment (FIG. 4). In addition to these transcription mediators (BCL11A and NF-E4), our data also showed that the phosphorylation level of the p38 was significantly increased by S51021 and SS-2394 (FIG. 5). Moreover, the pre-treatment of p38 MAPK inhibitor SB203580 efficiently abolished the elevation of globin gene expression triggered by S51021 and SS-2394 (data not shown). Taken together, it is demonstrated here that re-activation of the globin gene expression mediated by S51021 or SS-2394 is a complicated process, in which the modulations of transcription regulators (BCL11A and NF-E4) as well as p38 signaling participate in the regulatory program.

Cell Stress Response Involves in the Elevation of the Globin Gene Expression Mediated by HbF-Induing Agents.

[0272] Previous studies have demonstrated that 1,8-naphthalimides, such as amonafide, function as DNA intercalators and topoisomerase II (topo II) inhibitors in clinical development for the treatment of cancers including acute myeloid leukemia and prostate carcinoma. Norton et al., Anticancer Drugs. 2008; 19(1):23-36; and Allen et al., Expert Opinion on Investigational Drugs. 2011; 20(7):995-1003. The newly identified compounds, B1, B4, E2, and E3, which induce HbF, were previous found to exert their anti-cancer ability due to inhibition of topo II (DNA intercalator). Norton et al., Anticancer Drugs. 2008; 19(1):23-36; Stevenson et al., J Med Chem. 1984; 27(12):1677-1682; and Zee-Cheng et al., J Med Chem. 1985; 28(9):1216-1222. Considering the fact that cell growth inhibition and DNA intercalating cause cell stress response, it raises the possibility that cell stress response signaling may contribute to the elevation of globin gene expression. Indeed, several cell stress response-inducing agents, such as HU, thalidomide, TSA, and anisomycin, have been reported to activate the globin gene expression. Pace B et al., Exp Hematol. 2003; 31(11):1089-1096; Mabaera R et al, Exp Hematol. 2008; 36(9):1057-1072; Aerbajinai et al., Blood. 2007; 110(8):2864-2871; and Cokic V P., Journal of Clinical Investigation. 2003; 111(2):231-239. It also demonstrated that most of these cell stress response-inducing agents also inhibit cell proliferation and activate the p38 MAPK signaling pathway..sup.31 As shown in FIG. 5, both S51021 and SS-2394 inhibited cell growth and elevated the phosphorylation status of p38 MAPK at the concentrations of their IC.sub.50, suggesting that the cell stress response may be involved in the globin gene re-activation mediated by S51021 and SS-2394. However, the correlations between cell stress response and modulations of transcription regulators mediated by compound treatment need to be further investigated.

SS-2394 is a Lead Compound for Further Developing Novel HbF-Inducing Agent.

[0273] A a series of naphthalimide derivatives were synthesized for evaluating their HbF-inducing abilities. These agents were identified via lead optimization by structural modification of S51021. Of these derivatives, SS-2394 was found to have significant effect on increasing elevation of the globin mRNA. Although SS-2394 displayed lower IC.sub.50/EC ratio than that of S51021, several lines of evidence suggest that SS-2394 is a better lead compound for further drug development as an effective agent for treating P-thalassemia and sickle cell disease; First, SS-2394 shows higher globin gene induction fold at the concentration of its IC.sub.50 than that of S51021, suggesting that SS-2394 is a more efficient HbF-inducing agent than S51021 or others (FIG. 4A). Second, SS-2394 contains a symmetric pharmacophore which can be easily synthesized and purified. Third, this agent has greatly improved water-solubility and bioavailability and is more proper for new drug development (Table I). Fourth, SS-2394 displays higher HbF-inducing specificity than other compounds as demonstrated by the observation of the highest proportion of the globin mRNA (Table II). In sum, SS-2394, which is not a histone deacetylase inhibitor, was identified as a lead compound for further developing novel agents for treating hemoglobinopathies such as 3-thalassemia and sickle cell disease.

Example 2: Exemplary Compounds for Treating -Thalassemia and Sickle-Cell Disease

[0274] SS-2394 shows higher water-solubility than that of the parental compound S51021 but SS-2394 has much higher cytotoxicity than that of S51021 as evident by comparison of their IC.sub.50. To optimize the biological activity and reduce the cytotoxicity of SS-2394 by structural modification, the core structure of naphthalimide (I) was modified by alternating the carbonyl function of benzo[de]isoquinoline-1,3(2H)-dione; reducing the carbonyl function to hydroxyl derivatives (II), which can be further converted into dehydroxylated derivatives III and IV (Scheme 1). During biological evaluation of SS-2394, it was observed that the C6-amino function in SS-2394 was unstable and can be deaminated or converted into the corresponding C6-hydroxy derivatives. Therefore, various substituent R to the C6 of benzo[de]isoquinolin ring were introduced. The substituent R can be H, NH.sub.2, NHCOCH.sub.3, NHR, NR.sub.2; wherein R can be a C1 to C5 alkyl group. The new compounds bear a N,N-dimethylaminoethyl hydrophilic side chain, which can form acid salts with various inorganic acids or organic acids. Thus, the compounds are water soluble compounds and are suitable for biological application.

##STR00102##

General Methods

[0275] Several representative analogues which have been synthesized are provided in Table 2. The synthetic routes of these derivatives are displayed in Schemes 2, 3 and 4. See also Daffy et al., ChemistryA European Journal, 1998, 4, 1810-1815; Cheng; Journal of Medicinal Chemistry, 1985, 28, 1216-1222; and Lucatello et al., Bioorganic and Medicinal Chemistry, 2007, 15, 555-562.

[0276] Briefly, compound SS-2394 (1) was treated with acetic acid/acetic anhydride to give the N-acetyl derivatives (2, BO-2559), which was converted into 2,3-dihydro-1H-benzo[de]isoquinoline 3 (BO-2566) (Scheme 2). Treatment of 3 with HCl in EtOH gave the 6-amino-2,3-dihydro-1H-benzo[de]isoquinoline (4, BO-2560).

##STR00103##

[0277] To prevent the hydrolysis of C6-NH.sub.2 function, a NMe.sub.2 function was introduced to the C6 position of 2,3-dihydro-1H-benzo[de]isoquinoline-1,3(2H)-dione. Thus, compound 6-NMe.sub.2 substituted derivative BO-2561 (6, Scheme 3) was synthesized from C6-Cl derivative 5 by treating with dimethylamine in DMF in the presence of KOH. Under such reaction conditions, we also obtained C6-OH derivative (7, BO-2562) as the by-product. By following a similar procedure, compound 6 (BO-2561) was converted into 6-NH.sub.2-2,3-dihydro-1H-benzo[de]isoquinoline (8, BO-2563). The C6-NEt derivatives (9, BO-2564) was also synthesized by treating compound 5 with ethylamine. Similarly, compound 9 was further converted into the corresponding benzo[de]isoquinoline (10, BO-2565).

##STR00104##

[0278] Compound lacking one or two carbonyl function(s) were synthesized for evaluating their biological activity. As shown in Scheme 4, dione 11 was treated with NaBH.sub.4 in ethanol to give mono-hydroxy derivative 12 (BO-2476), which was further converted into mono-carbonyl derivative 13 (BO-2477). Similarly, dione 11 was converted into the corresponding benzo[de]isoquinoline 14 (BO-2563).

##STR00105##

Compounds

N-(2-(2-(Dimethylamino)ethyl)-1,3-dioxo-2,3-dihydro-1H-benzo[de]isoquinolin-6-yl)acetamide (2, BO-2559)

[0279] Daffy et al., ChemistryA European Journal, 1998, 4, 1810-1815. A suspension of known 6-amino-2-(2-(dimethylamino)-ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (1, 640 mg, 2 mmol) in a mixture of acetic acid (24 mL) and acetic anhydride (3 mL) was refluxed for 45 min. Methanol was added to destroy excess acetic anhydride and then co-evaporated with methanol several times to dryness. The yellow product was collected by filtration to give the desired product N-(2-(2-(dimethylamino)ethyl)-1,3-dioxo-2,3-dihydro-1H-benzo-[de]isoquinolin-6-yl)acetamide (2, BO-2559),.sup.1 590 mg (90%); mp. 278-279 C.; .sup.1H NMR (DMSO-d.sub.6) 2.30 (3H, s, CH.sub.3), 2.87 (6H, s, N(CH.sub.3).sub.2), 3.41 (2H, t, J=5.9 Hz, CH.sub.2), 4.38 (2H, t, J=5.9 Hz CH.sub.2), 7.91 (1H, t, J=8.5 Hz, ArH), 8.34 (1H, d, J=8.2 Hz, ArH), 8.50 (1H, d, J=8.2 Hz, ArH), 8.55 (1H, d, J=6.6 Hz, ArH), 8.79 (1H, d, J=8.6 Hz, ArH), 10.52 (1H, br, NH). HRMS [ES.sup.+]: calcd for C.sub.18H.sub.19N.sub.3O.sub.3, 325.3618 [M+H].sup.+, found 326.1500.

2-(2-(Dimethylamino)ethyl)-2,3-dihydro-1H-benzo[de]isoquinolin-6-amine (4, BO-2560)

[0280] LiAlH.sub.4 (20 mg, 3 mmol) was added portionwise to a stirring suspension of AlCl.sub.3 (85 mg, 0.62 mmol) in dry THF (10 mL) in an ice-water bath under Argon and stirred for 10 min. N-(2-(2-(dimethylamino)ethyl)-1,3-dioxo-2,3-dihydro-1H-benzo[de]isoquinolin-6-yl)acetamide (2, 163 mg, 0.5 mmol) was added portionwise to the above suspension. The reaction mixture was stirred at room temperature for 20 min and then heated to 40 C. for 1 hr. The reaction mixture was poured into ice-water and the solid was removed by filtration through a pad of Celite. The filtrates were evaporated to dryness under reduced pressure and the residue was dissolved in ethanol (50 mL) and then HCl.sub.(conc) (0.5 mL) and then evaporated to dryness. The solid product was recrystallized from ethanol to give 2-(2-(dimethylamino)ethyl)-2,3-dihydro-1H-benzo[de]isoquinolin-6-amine (4, BO-2560), 101 mg (78%); mp. 253-254 C.; .sup.1H NMR (DMSO-d.sub.6) 2.86 (6H, s, N(CH.sub.3).sub.2), 3.70 (4H, s, 2CH.sub.2), 4.65-4.97 (4H, s, 2CH.sub.2), 6.65 (1H, d, J=7.8 Hz, ArH), 7.28 (1H, d, J=7.8 Hz, ArH), 7.42 (1H, d, J=7.1 Hz, ArH), 7.49 (1H, t, J=7.8 Hz, ArH), 8.22 (1H, d, J=8.3 Hz, ArH), 10.83 (2H, br, NH.sub.2). HRMS [ES.sup.+]: calcd for C.sub.16H.sub.21N.sub.3, 255.3580 [M+H].sup.+, found 256.1814.

2-(2-(dimethylamino)ethyl)-6-hydroxy-H-benzo[de]isoquinoline-1,3(2H)-dione (6, BO-2562).SUP.2 .and 6-(dimethylamino)-2-(2-(dimethylamino)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (7, BO-2561)

[0281] A solution of 6-chloro-2-(2-(dimethylamino)ethyl)-1H-benzo-[de]isoquinoline-1,3(2H)-dione (5, 1.5 g, 5 mmol) and KOH (1.12 g, 20 mmol) in 10 mL of DMF was heated at reflux for 45 min. The solvent was then evaporated in vacuo to dryness. The residue was chromotographered on a silica gel column (420 cm) using CHCl.sub.3:MeOH (100:2 v/v) as the eluent. The first main product eluated was collected to give 6-(dimethylamino)-2-(2-(dimethyl-amino)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (7, BO-2561), 795 mg (50%), mp. 198-199 C. .sup.1H NMR (DMSO-d.sub.6) 2.89 (6H, s, N(CH.sub.3).sub.2), 3.12 (6H, s, N(CH.sub.3).sub.2), 3.44 (2H, t, J=5.9 Hz, CH.sub.2), 4.38 (2H, t, J=5.9 Hz, CH.sub.2), 7.24 (1H, d, J=7.8 Hz, ArH), 7.79 (1H, t, J=7.9 Hz, ArH), 8.37 (1H, d, J=8.4 Hz, ArH), 8.50 (1H, d, J=7.2 Hz, ArH), 8.56 (1H, d, J=8.4 Hz, ArH). HRMS [ES.sup.+]: calcd for C.sub.18H.sub.21N.sub.3O.sub.2, 311.3782 [M+H].sup.+, found 312.1061).

[0282] The second product eluated was collected to give 2-(2-(dimethylamino)ethyl)-6-hydroxy-1H-benzo[de]isoquinoline-1,3(2H)-dione (6) (Daffy et al., ChemistryA European Journal, 1998, 4, 1810-1815) 160 mg (11%); mp. 202-204 C.; .sup.1H NMR (DMSO-d.sub.6) 2.38 (6H, s, N(CH.sub.3).sub.2), 2.71 (2H, t, J=8.3 Hz, CH.sub.2), 4.19 (2H, t, J=8.3 Hz, CH.sub.2), 6.77 (1H, d, J=8.5 Hz, ArH), 7.56 (1H, t, J=7.7 Hz, ArH), 8.18 (1H, d, J=8.5 Hz, ArH), 8.37 (1H, d, J=7.2 Hz, ArH), 8.48 (1H, d, J=8.1 Hz, ArH). HRMS [ES.sup.+]: calcd for C.sub.16H.sub.16N.sub.2O.sub.3, 284.3098 [M+H].sup.+, found 285.1240.

2-(2-(Dimethylamino)ethyl)-N,N-dimethyl-2,3-dihydro-1H-benzo[de]isoquinolin-6-amine (8, BO-2563)

[0283] LiAlH.sub.4 (20 mg, 3 mmol) was added portionwise to a stirring suspension of AlCl.sub.3 (85 mg, 0.62 mmol) in 10 mL of THF in an ice-water bath under Argon for 10 min. 6-(dimethylamino)-2-(2-(dimethylamino)ethyl)-1H-benzo[de]-isoquinoline-1,3(2H)-dione (7, 311 mg, 1 mmol) was added portionwise. The reaction mixture was allowed to stir at room temperature and then heated to 40 C. for 1 h. The reaction mixture was poured into ice-water and filtered through a pad of Celite. The filtrate were evaporated to dryness under reduced pressure. The solid product was recrystallized from EtOH to give 2-(2-(dimethylamino)ethyl)-N,N-dimethyl-2,3-dihydro-1H-benzo[de]isoquinolin-6-amine (8, BO-2563), 184 mg (65%), mp. 251-252 C.; .sup.1H NMR (DMSO-d.sub.6) 2.89 (6H, s, N(CH.sub.3).sub.2), 2.95 (6H, s, N(CH.sub.3).sub.2), 3.94 (4H, S, 2CH.sub.2), 4.71 (2H, S, CH.sub.2), 4.76 (2H, S, CH.sub.2), 7.36 (1H, d, J=7.2 Hz, ArH), 7.46 (1H, d, J=7.8 Hz, ArH), 7.52 (1H, d, J=7.2 Hz, ArH), 7.66 (1H, t, J=7.8 Hz, ArH), 8.19 (1H, d, J=8.5 Hz, ArH). HRMS [ES.sup.+]: cald for C.sub.18H.sub.25N.sub.3, 283.4112 [M+H].sup.+, found 284.2129.

2-(2-(Dimethylamino)ethyl)-6-(ethylamino)-1H-benzo[de]isoquinoline-1,3(2H)-dione (9, BO-2564)

[0284] A solution of 6-chloro-2-(2-(dimethylamino)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (1.8 g, 6 mmol), and KOH (1.68 g, 30 mmol) in 4 mL of N,N-diethylformamide was refluxed for 2 hr. The solvent was evaporated to dryness. The residue was chromotographer on silica gel (620 cm) using CHCl.sub.3:methanol 100:4 as eluant to give syrup product 2-(2-(dimethylamino)ethyl)-6-(ethylamino)-1H-benzo[de]isoquinoline-1,3(2H)-dione (9, BO-2564), 313 mg (10%); mp. 253-4 C.; .sup.1H NMR (DMSO-d.sub.6) 1.30 (3H, t, J=7.1 Hz, CH.sub.2CH.sub.3), 2.20 (6H, s, N(CH.sub.3).sub.2), 2.48. (2H, t, J=6.9 Hz, CH.sub.2), 3.42 (2H, q, J=7.1 Hz, CH.sub.2CH.sub.3) 4.13 (2H, t, J=6.9 Hz, CH.sub.2), 6.79 (1H, d, J=8.5 Hz, ArH), 7.68 (1H, t, J=7.9 Hz, ArH), 7.74 (1H, br, NH), 8.28 (1H, d, J=8.5 Hz, ArH), 8.42 (1H, d, J=9.0 Hz, ArH), 8.44 (1H, d, J=7.3 Hz, ArH), 8.69 (1H, d, J=8.5 Hz, ArH). HRMS [ES.sup.+]: calcd for C.sub.18H.sub.21N.sub.3O.sub.2, 311.3782 [M+H].sup.+, found 312.1814.

2-(2-(dimethylamino)ethyl)-N-ethyl-2,3-dihydro-1H-benzo[de]isoquinolin-6-amine (10, BO-2565)

[0285] A suspension of AlCl.sub.3 (85 mg, 0.62 mmol) in 10 mL of THF was added portionwise LiAlH.sub.4 (20 mg, 3 mmol) at ice-water bath under Argon for 10 min. 2-(2-(dimethylamino)ethyl)-6-(ethylamino)-1H-benzo[de]isoquinoline-1,3 (2H)-dione (170 mg, 0.5 mmol) was added portionwise at ice-water. It was stirred at room temperature and then heated to 40 C. for 1 hr. The reaction mixture was poured into ice-water. The solid was removed by a pad of Celite. The filtrates were evaporated to dryness. The product was recrystallized from EtOH/HCl to give 2-(2-(dimethyl-amino)ethyl)-N-ethyl-2,3-dihydro-1H-benzo[de]isoquinolin-6-amine, 38 mg (24%), mp. 239-240 C.; .sup.1H NMR (DMSO-d.sub.6) 1.30 (3H, t, J=7.0 Hz, CH.sub.2CH.sub.3), 2.87 (6H, s, N(CH.sub.3).sub.2), 3.27 (2H, q, J=7.0 Hz, CH.sub.2CH.sub.3), 3.69 (4H, brs, 2CH.sub.2), 4.87 (4H, brs, 2CH.sub.2), 6.61 (1H, d, J=7.3 Hz, ArH), 7.29 (1H, d, J=7.7 Hz, ArH), 7.49 (1H, br, NH), 7.62 (1H, t, J=7.7 Hz, ArH), 7.98 (1H, d, J=8.2 Hz, ArH), 8.20 (1H, d, J=8.4 Hz, ArH). HRMS [ES.sup.+]: calcd for C.sub.18H.sub.25N.sub.3, 283.4112 [M+H].sup.+, found 284.3225.

2-(2-(Dimethylamino)ethyl)-3-hydroxy-2,3-dihydro-1H-benzo[de]isoquinolin-1-one (12, BO-2476)

[0286] NaBH.sub.4 (0.37 g, 10 mmol) was added portionwise to a solution of 2-(2-(dimethylamino)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (11, 0.27 g, 1 mmol) in EtOH/Water (20 mL, v/v:10/1). The mixture was stirred overnight at room temperature. After the completion of the reaction, the reaction mixture was evaporated under reduced pressure to dryness. The solid residue was triturated with water and extracted with CH.sub.2Cl.sub.2 (100 mL3). The combined organic extracts were dried over Na.sub.2SO.sub.4 and evaporated to dryness and the solid was recrystallized from Haxen/CH.sub.2Cl.sub.2 to afford the desired product 2-(2-(dimethylamino)ethyl)-3-hydroxy-2,3-dihydro-1H-benzo[de]isoquinolin-1-one (12, BO-2476), 0.24 g (89%); mp. 129-130 C.; .sup.1H NMR (DMSO-d.sub.6) 2.26 (6H, s, N-Me.sub.2), 2.48-2.45 (1H, m, CH), 2.72-2.68 (1H, m, CH), 3.71-3.65 (1H, m, CH), 3.97-3.92 (1H, m, CH), 6.15 (1H, s, CHOH), 7.71-7.64 (3H, m, ArH), 7.78 (1H, s, CHOH, exchangeable), 8.03 (1H, d, J=7.1 Hz, ArH), 8.19 (1H, d, J=8.2 Hz, ArH), 8.24 1H, d, J=7.1 Hz, ArH). HRMS [ES.sup.+]: cald for C.sub.16H.sub.18N.sub.2O.sub.2, 270.3262 [M+H].sup.+, found 271.2231.

2-(2-(Dimethylamino)ethyl)-2,3-dihydro-1H-benzo[de]isoquinolin-1-one (13, BO-2477)

[0287] To a solution of 2-(2-(dimethylamino)ethyl)-3-hydroxy-2,3-dihydro-1H-benzo[de]isoquinolin-1-one (12, 0.27 g, 1 mmol) in CH.sub.2Cl.sub.2 (10 mL) were added triethylsilane (0.48 mL, 3 mmol) and TFA (4 mL). The reaction mixture was stirred overnight at room temperature. After the completion of, solvent was evaporated under reduced pressure to dryness and the residue was triturated with saturated NaHCO.sub.3 solution and then extracted with CH.sub.2Cl.sub.2. The combined organic extracts were dried over Na.sub.2SO.sub.4 and evaporated to dryness. The solid residue was recrystallized from Haxen/CH.sub.2Cl.sub.2 to 2-(2-(dimethylamino)ethyl)-2,3-dihydro-1H-benzo[de]isoquinolin-1-one (13, BO-2477), 0.22 g, 86%; mp. 253-254 C.; .sup.1H NMR (DMSO-d.sub.6) 2.86 (6H, s, N-Me.sub.2), 3.48-3.45 (2H, m, CH.sub.2), 4.02-3.99 (2H, m, CH.sub.2), 5.14 (2H, s, CH.sub.2), 7.48 (1H, d, J=7.0 Hz, ArH), 7.66-7.59 (2H, m, ArH), 7.91 (1H, d, J=8.2 Hz, ArH), 8.16-8.12 (2H, m, ArH). HRMS [ES.sup.+]: cald for C.sub.16H.sub.18N.sub.2O, 254.3269 [M+H].sup.+, found 255.1169.

2-(1H-Benzo[de]isoquinolin-2(3H)-yl)-N,N-dimethylethan-1-amine (14, BO-2478)

[0288] Lucatello et al., Bioorganic and Medicinal Chemistry, 2007, 15, 555-562. To suspension of AlCl.sub.3 (0.17 g, 1.25 mmol) in anhydrous THF was added LiAlH.sub.4 (0.24 g, 6 mmol) followed by slow addition of 2-(2-(dimethyl-amino)ethyl)-1H-benzo[de]isoquinoline-1,3(2H)-dione (11, 0.27 g, 1 mmol) at 0 C. The mixture was then refluxed for 3 h, cooled to 0 C. and then poured into ice/THF cautiously. The solid separated was collected by filtration and the filtrate was extracted with CH.sub.2Cl.sub.2. The organic layer was separated and dried over anhydrous Na.sub.2SO.sub.4 and evaporated to dryness afforded the final product. The final product was then converted in to HCl salt by treating with EA/HCl to give 2-(1H-benzo[de]isoquinolin-2(3H)-yl)-N,N-dimethylethan-1-amine (14, BO-2478), 0.215 g (78%); mp. 208-209 C.; .sup.1H NMR (CDCl.sub.3-d.sub.6) 2.28 (6H, s, N(CH.sub.3).sub.2), 2.62-2.58 (2H, m, CH.sub.2), 2.78-2.75 (2H, m, CH.sub.2), 4.02 (2H, s, CH.sub.2), 7.19 (2H, d, J=8.6 Hz, ArH), 7.41-7.36 (2H, m, ArH), 7.68 (2H, d, J=10.3 Hz, ArH). HRMS [ES.sup.+]: cald for C.sub.16H.sub.21N.sub.2Cl, 276.8043 [M+H].sup.+, found 277.7241.

Biological Activity

[0289] All the newly synthesized compounds were evaluated for their globin-inducing abilities. FIG. 8 shows the effect of globin-induction and cytotoxicity against primary erythroid cells by the representative analogues of 2,3-dihydro-1H-benzo[de]isoquinoline (line 1: mock control; line 2: hydroxyurea (HU); line 3: sodium butyrate (NaB); line 4: SS-2394; line 5: S51021; line 6: BO-2562; line 7: BO-2561; line 8: BO-2559; line 9: BO-2566; line 10: BO-2563, line 11: BO-2560; line 12: BO-2565; line 13; BO-2477; line 14: BO-2476; line 15: BO-2478. The tested concentrations of each compound and the survival rates of the primary erythroid cells after 2 days of compound treatment are listed at the bottom of the graph.

[0290] As shown in FIG. 8, the newly synthesized compounds BO-2559, BO-2560, BO-2561, BO-2476 and BO-2477 showed significant globin-inducing abilities, of which the expression levels of globin gene were up-regulated by 2.1-6.9 folds in primary erythroid cells. All these newly identified globin-inducing compounds have reduced cytotoxicities with IC.sub.50 ranging from 5 to 40 M against primary erythroid cells. Among the compounds tested, compound BO-2477 was shown to have the highest globin-inducing activity with relatively lower cytotoxicity.