Deuterated retinoid compounds and their synthesis and uses

Abstract

The present disclosure relates to novel deuterated retinoid compounds and methods for preparing the deuterated retinoid compounds. The present disclosure also provides pharmaceutical compositions comprising the deuterated retinoid compounds, and uses of the deuterated retinoid compounds for treating diseases, including breast cancer and prostate cancer.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is based on and claims priority to U.S. Provisional Application No. 63 / 316,328, filed March 3, 2022, and U.S. Provisional Application No. 63 / 487,705, filed March 1, 2023, the entire contents of which are incorporated herein by reference.

[0002] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT This invention was made with Government support under Grant Nos. CA129379 and CA224696 awarded by the National Institutes of Health. The Government has certain rights in this invention.

[0003] Technical Field The present disclosure generally relates to novel deuterated retinoid compounds useful as degradation inducers of mitogen-activated protein kinase (MAPK) interacting kinases 1 and 2 (Mnk1 / 2). The present disclosure also provides methods for preparing the deuterated retinoid compounds, pharmaceutical compositions comprising the deuterated retinoid compounds, and methods of using the deuterated retinoid compounds in the treatment of various Mnk1 / 2-related diseases. [Background technology]

[0004] There continues to be a need for highly effective therapeutic agents for treating cancer with a sufficiently large therapeutic window and no or limited toxicity and side effects. The present inventors have discovered novel deuterated retinoid compounds with unique properties and biological activities, including modulation of the Mnk1 / 2-eukaryotic translation initiation factor (eIF4E) and androgen receptor (AR) signaling pathways, that can treat a variety of cancers and diseases that depend on functional Mnk1 / 2, androgen receptor, and / or androgen receptor splice variants for their pathogenesis while limiting adverse side effects. Summary of the Invention

[0005] The present disclosure relates to a compound of formula (A)

[0006] [ka] or a pharma- ceutically acceptable salt thereof, 1 ~R 8 Each of may be independently H or D, and R 1 ~R 8 wherein at least one of R is D. 4 , and R 6 ~R 8 At least one of may be D.

[0007] In some embodiments, the compound is

[0008] [ka] [ka] [ka] or a pharma- ceutically acceptable salt thereof. In some embodiments, the compound is

[0009] [ka] [ka] or a pharma- ceutically acceptable salt thereof.

[0010] The present disclosure also provides pharmaceutical compositions.The pharmaceutical compositions can include the above-mentioned compounds or their pharmaceutically acceptable salts and pharmaceutically acceptable carriers.In some embodiments, the pharmaceutical compositions can be formulated for oral administration.

[0011] The present disclosure also provides a method for treating disease.The method can include administering a therapeutically effective amount of the above-mentioned compound or its pharma-ceutically acceptable salt to a subject in need thereof.In some embodiments, the disease can be one of breast cancer, prostate cancer, bladder cancer, pancreatic cancer, hepatocellular carcinoma, benign prostatic hyperplasia, Kennedy's disease, and blood cancer.In some embodiments, the compound is administered orally.

[0012] The present disclosure also relates to a compound of formula (A):

[0013] [ka] or a salt thereof. The method comprises reacting a compound of formula (B):

[0014] [ka] or a salt thereof, 1 ~R 8 is independently H or D, and R 1 ~R 8 at least one of is D.

[0015] In some embodiments, the conversion of the compound of formula (B) comprises converting the compound of formula (B) to a compound of formula (C) or a salt thereof or a compound of formula (D) or a salt thereof:

[0016] [ka] This may include converting the

[0017] In some embodiments, the method comprises reacting a compound of formula (C) or a salt thereof with a compound of formula (E) or a salt thereof or a compound of formula (F) or a salt thereof:

[0018] [ka] The method may further include converting the

[0019] In some embodiments, the method comprises reacting a compound of formula (D) or a salt thereof with a compound of formula (G) or a salt thereof or a compound of formula (H) or a salt thereof:

[0020] [ka] The method may further include converting the

[0021] The present disclosure also relates to a compound of formula (I)

[0022] [ka] or a salt thereof. The method comprises the steps of:

[0023] [ka] to a compound of formula (I).

[0024] In some embodiments, the conversion of the compound of formula (B) is Reacting a compound of formula (B) with a compound of formula (C):

[0025] [ka] Converting to Reacting a compound of formula (C) with a compound of formula (E):

[0026] [ka] Then convert it to Converting a compound of formula (E) to a compound of formula (I) may include.

[0027] The subject matter which is regarded as the invention is particularly pointed out and distinctly claimed in the concluding portion of the specification. The objects, features, and advantages of the present disclosure will become apparent from the following detailed description taken in conjunction with the accompanying drawings. [Brief description of the drawings]

[0028] [Figure 1] FIG. 1 shows the chemical structures of all-trans retinoic acid (ATRA) (Compound 1) and the development candidate (±)-4-imidazolyl-4(H)-(4'-fluoro(phenyl))-(E)-retinamide (VNLG-152R, Compound 2). [Diagram 2] FIG. 1 shows predicted metabolic sites on VNLG-152R (compound 2). Atoms of VNLG-152R most likely for oxidation by Cyp3A4, Cyp2D6 and Cyp2C9 are marked with circles (◯), while other predicted metabolic sites are marked with squares (□) and triangles (△), representing amide bond cleavage and aromatic ring hydroxylation to form phenols, respectively. [Diagram 3] FIG. 1 shows the chemical structures, molecular formulas (MF), and accurate masses (EM) of VNLG-152R (compound 3) and its deuterated analogues (compounds 3-9). [Figure 4] FIG. 1 shows the synthesis of VNLG-152R from ATRA according to Scheme 1 of the present disclosure. [Diagram 5] FIG. 1 shows the synthesis of VNLG-152R from 4-oxo-ATRA (compound 15) according to Scheme 2 of the present disclosure. [Figure 6] FIG. 1 shows the synthesis of deuterated retinoid compounds, compounds 3 to 5, of the present disclosure, according to Scheme 3 of the present disclosure. [Figure 7] FIG. 1 shows the synthesis of deuterated retinoid compounds, compounds 6-9 of the present disclosure, according to Scheme 4 of the present disclosure. [Figure 8A] FIG. 8A shows a direct comparison of the in vitro anti-proliferative effects of non-deuterated VNLG-152R and its deuterated analogs, compounds 3-9 of the present disclosure, on MDA-MB-231 (FIG. 8A) human triple-negative breast cancer cells. [Figure 8B] FIG. 8B shows a direct comparison of the in vitro anti-proliferative effects of non-deuterated VNLG-152R and its deuterated analogs, compounds 3-9 of the present disclosure, on MDA-MB-468 (FIG. 8B) human triple-negative breast cancer cells. [Figure 9A] FIG. 9A shows a direct comparison of the in vitro anti-proliferative effects of non-deuterated VNLG-152R and its deuterated analogs, compounds 3-9 of the present disclosure, and paclitaxel (PTX) on MDA-MB0231 (FIG. 9A) human triple-negative breast cancer cells. [Figure 9B] FIG. 9B shows a direct comparison of the in vitro anti-proliferative effects of non-deuterated VNLG-152R and its deuterated analogs, compounds 3-9 of the present disclosure, and paclitaxel (PTX) on MDA-MB-468 (FIG. 9B) human triple-negative breast cancer cells. [Figure 10] FIG. 1 is a schematic diagram of Mnk1 / 2-eIF4E signaling and the action sites of the disclosed Mnk1 / 2 degradation inducers. [Figure 11A] FIG. 11A shows the effect of VNLG-152R and its deuterated analogs, compounds 3-9 of the present disclosure, on in vitro expression of Mnk1, peIF4E, eIF4E, BCL2, BAX, and cyclin D1 in MDA-MB-231 (FIG. 11A) human triple-negative breast cancer cells. [Figure 11B] FIG. 11B shows the effect of VNLG-152R and its deuterated analogs, compounds 3-9 of the present disclosure, on the in vitro expression of Mnk1, peIF4E, eIF4E, BCL2, BAX, and cyclin D1 in MDA-MB-468 (FIG. 11B) human triple-negative breast cancer cells. [Figure 12A] Figure 12 shows the plasma pharmacokinetic profiles of single individual or cassette PO administration of VNLG-152R and compounds 3-9 of the present disclosure to CD-1 mice. Data are mean (±SD) concentrations from three CD-1 mice. Figure 12A represents cassette administration. [Figure 12B]Figure 12B shows the plasma pharmacokinetic profiles of single individual or cassette PO administration of VNLG-152R and compounds 3-9 of the present disclosure to CD-1 mice. Data are mean (±SD) concentrations from three CD-1 mice. Figure 12B represents cassette administration. [Figure 12C] Figure 12 shows the plasma pharmacokinetic profiles of single individual or cassette PO administration of VNLG-152R and compounds 3-9 of the present disclosure to CD-1 mice. Data are mean (±SD) concentrations from three CD-1 mice. Figure 12C represents individual administration. [Figure 13A] Figure 13 shows the antitumor activity of VNLG-152R, Compound 4, Compound 8, and Compound 9 in xenograft mouse models. Figure 13A shows tumor size after 16 days of treatment (PO, 20 mg / kg, 5 days / week) in the MDA-MB-231 model. [Figure 13B] Figure 13 shows the antitumor activity of VNLG-152R, compound 4, compound 8, and compound 9 in xenograft mouse model. Figure 13B shows the percentage change in tumor volume relative to the volume at the end of the experiment for MDA-MB-231 tumor xenografts. In Figure 13B, *=p<0.0001. [Figure 13C] Figure 13 shows the antitumor activity of VNLG-152R, Compound 4, Compound 8, and Compound 9 in a xenograft mouse model. Figure 13C shows tumor size after 16 days of treatment (PO, 20 mg / kg, 5 days / week) in the MDA-MB-468 model. [Figure 13D] Figure 13 shows the antitumor activity of VNLG-152R, compound 4, compound 8, and compound 9 in xenograft mouse model. Figure 13D shows the percentage change in tumor volume relative to the volume at the end of the experiment for MDA-MB-468 tumor xenografts. In Figure 13D, *=p<0.0001. [Figure 14A] Figure 14 shows the in vitro effects of VNLG-152R on Synoviolin 1 (SYVN1) and other targets in MDA-MB-231 cells. Figure 14A shows that 20 μM VNLG-152R induces SYVN1. [Figure 14B] Figure 14B shows the in vitro effects of VNLG-152R on Synoviolin 1 (SYVN1) and other targets in MDA-MB-231 cells. Figure 14B shows that VNLG-152R induces SYVN1 in a dose-dependent manner and suppresses the levels of Mnk1 and downstream targets. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0029] cancer Breast cancer is the second most common cancer among women in the United States, after certain types of skin cancer (Centers for Disease Control and Prevention, Office of Cancer Prevention and Control, Breast Cancer Statistics, available at https: / / www.cdc.gov / cancer / breast / statistics / index.htm (last accessed February 15, 2022)). Although early diagnosis and new treatment options have been effective in managing primary breast cancer, treatment of advanced breast cancer, especially metastatic disease, remains a challenge. 1 Triple-negative breast cancer (TNBC) lacks expression of estrogen receptor (ERα), progesterone receptor (PgR), and human epidermal growth factor receptor 2 (HER2). TNBC is also the leading cause of breast cancer mortality due to its aggressive invasive and metastatic capabilities, lack of suitable molecular therapeutic targets, and resistance to conventional chemotherapy agents. 2 Therefore, there is an urgent need to identify and develop new therapeutic agents that are effective against TNBC and metastatic breast cancer (MBC), especially those that can provide higher survival rates, fewer side effects, and a better quality of life for patients than currently available therapeutic agents.

[0030] Meanwhile, prostate cancer is one of the most common malignancies among men worldwide. 3Androgen deprivation therapy is typically used to treat advanced and metastatic prostate cancer. However, in most cases, prostate cancer eventually becomes unresponsive to androgen deprivation and develops into castration-resistant prostate cancer (CRPC), which then progresses rapidly. Treatment of CRPC remains limited, with only a few drugs providing reasonable survival in patients with metastatic CRPC (mCRPC). 4 Thus, as in breast cancers such as TNBC and MBC, there is an urgent need for new effective therapeutic agents for prostate cancers such as mCRPC with potential antitumor activity and low toxicity.

[0031] Eukaryotic translation initiation factor 4E (eIF4E), a component of the "eIF4F cap-binding complex," plays a key role in mRNA discrimination and driving the development and progression of various cancers, including breast and prostate cancer. eIF4G-associated eIF4E binds to the m7G cap at the 5' end of eukaryotic mRNAs and initiates the translation of "weak mRNAs" that code for malignancy-associated proteins. 5、6 The oncogenic potential of eIF4E is dependent on its phosphorylation at serine 209 by Mnk1 / 2, and importantly, eIF4E(S209) phosphorylation by Mnk1 / 2 is critical for tumorigenicity but not for normal mammalian development. 7~9 eIF4E knockdown has been shown to reduce breast cancer cell proliferation in rapamycin-sensitive and rapamycin-insensitive breast cancer cell lines. 10 eIF4E depletion has also been shown to enhance the antiproliferative and proapoptotic effects of chemotherapeutic drugs in breast cancer cells. 11 Treatment of human breast cancer cells with MNK inhibitors has been demonstrated to reduce colony formation, proliferation, and survival. 12The impact of protein translation has also been clearly demonstrated in clinical practice. For example, a recent prospective study concluded that over 60% of TNBC patients have tumors with high levels of eIF4E, TNBC patients with high eIF4E overexpression are more likely to relapse and die from cancer recurrence, and high eIF4E appears to be an important prognostic marker in TNBC patients. 13 eIF4E is involved in breast cancer progression and angiogenesis. 14 , and metastasis 15 A 7-fold or greater increase in eIF4E is a strong prognostic indicator of breast cancer recurrence and death in both retrospective and prospective clinical studies. 16~18 Moreover, recent reports have highlighted the impact of inhibition of Mnk-eIF4E signaling in experimental and clinical settings for TNBC. 19~21 .

[0032] Epithelial-mesenchymal transition (EMT) is a highly conserved process that allows polarized immobile epithelial cells to transdifferentiate into cells with a motile mesenchymal phenotype. 22、23 EMT produces cancer cells that exhibit stem cell properties, are invasive, migratory, and have metastatic potential. 24 EMT has been shown to play an important role in the development of mCRPC. 25、26 Full-length androgen receptor (f-AR) signaling and activation of the Mnk1 / 2-eIF4E pathway are known to promote EMT and contribute to metastasis by facilitating signaling interactions. 27~29 f-AR induces EMT through activation of Snail transcription factor or suppression of E-cadherin 27 Activation of Snail increases the expression of mesenchymal markers and invasion-related proteins. In contrast to activation of Snail, transcriptional repression of E-cadherin, a key mediator of intercellular adhesion at adherens junctions, leads to disruption of cell-cell communication and initiation of EMT. 30、31 .

[0033] Activation of eIF4E is recognized to stimulate EMT by promoting the expression of pro-metastatic factors such as Snail and matrix metalloproteinases (MMPs). 29 MMPs cleave several components of the extracellular matrix (ECM) to promote tissue invasion and metastasis. 32、33 In addition to supporting tumor initiation, activation of eIF4E has been demonstrated to promote metastatic progression through the translation of several EMT-associated mRNAs, such as those for Snail and MMP-3. 29 .

[0034] There is growing interest in developing drugs that inhibit or disrupt the oncogenic eIF4F translation complex (and, consequently, eIF4E) as a strategy to obtain effective anticancer therapeutics for a variety of solid tumors and hematological cancers. 34~44 Among the various strategies to disrupt the eIF4F complex, inhibition of Mnk1 / 2 to prevent eIF4E phosphorylation has attracted strong interest. The finding that Mnk1 and Mnk2 regulate mammalian target of rapamycin complex 1 (mTORC1) signaling and associate with mTORC1 directly suggests that Mnk1 / 2 inhibitors and degradation inducers can also inhibit mTORC1 / 4E-BP1 / p70SK6 signaling. 45、46 Thus, pharmacological inhibition of Mnk1 / 2, and therefore of both the Mnk-eIF4E and mTORC1 pathways, may serve as a potentially effective therapeutic approach for treating patients with advanced breast cancer, including TNBC and MBC, as well as other malignancies that likely have dysregulated Mnk-eIF4E / m-TORC1 signaling.

[0035] Retinoid Compounds Retinoic acid metabolism blockers (RAMBAs), a family of compounds that inhibit the P450 enzymes responsible for the metabolism of all-trans retinoic acid (ATRA), have been shown to exert potent anticancer and growth inhibitory effects in human breast and prostate cancer cells in vitro and in xenograft models in vivo.47~52 Retinoid compounds are a group of natural and synthetic analogues of ATRA and 4-hydroxyphenylretinamide (4-HPR). Retinoid compounds comprise a family of polyisoprenoid compounds and are currently the subject of intense biological interest, prompted by the discovery and characterization of retinoid receptors and the recognition of these compounds as nonsteroidal small molecule hormones. Retinoid compounds can inhibit proliferation and induce terminal differentiation and apoptosis in cultured cancer cell lines, leading to widespread interest in their use in cancer therapy. 53 The biological effects of retinoid compounds result from the regulation of gene expression mediated through two complex types of nuclear receptors, the retinoic acid receptors and the retinoid X receptors (RARs and RXRs). 54 Each type includes three distinct subtypes (α, β, and γ) that are encoded by different genes. Each RAR and RXR subtype is expressed in a specific pattern in different tissues and is thought to have a specific profile of gene regulatory activity. Nuclear receptors function as dimers. RARs form heterodimers with RXRs. RXRs are more versatile and bind to RARs as well as other nuclear receptors, including thyroid hormone receptors and vitamin D receptors.

[0036] Applicants have designed and synthesized a class of novel retinoid compounds that exert potent anticancer effects in breast and prostate cancer cells and tumor xenografts by inducing ubiquitin-proteasomal degradation of Mnk1 / 2 and preventing eIF4E and mTORC1 activation, thereby resulting in inhibition of cancer cell proliferation, apoptosis evasion, cell migration and invasion in vitro, and tumor xenograft inhibition in vivo. 44、47、48、55、56 One class of novel retinoid compounds is based on the ATRA scaffold (compound 1) (Figure 1). ATRA retinoid compounds are as described in U.S. Patent No. 7,265,143 and U.S. Patent No. 9,156,792, the contents of each of which are incorporated herein by reference in their entirety.

[0037] The applicants have demonstrated that a novel ATRA retinoid compound simultaneously targets both f-AR / androgen receptor splice variant-7 (AR-V7) signaling and MNK-eIF4E translation in several androgen-sensitive and CRPC cells in vitro by enhancing f-AR / AR-V7 and MNK degradation via the ubiquitin-proteasome pathway. 47 , and induced significant downregulation of f-AR / AR-7, MNK-1 / 2, and p-eIF4E, inhibiting tumor growth and development mediated by f-AR- and MNK-induced eIF4E activation in xenograft models of CRPC in vivo 55 Notable ATRA retinoid compounds are the lead racemic VNLG-152 (VNLG-152R) (compound 2, (2E,4E,6E,8E)-9-(3-imidazolyl-2,6,6-trimethylcyclohex-1-enyl)-3,7-dimethyl-N-phenylnona-2,4,6,8-tetraenamide), VNLG-153 (compound 22, (2E,4E,6E,8E)-9-(3-imidazolyl-2, The racemic VNLG-152, VNLG-153, and VNLG-147 (compound 23, (2E,4E,6E,8E)-N-(2-hydroxyphenyl)-9-(3-imidazolyl-2,6,6-trimethylcyclohex-1-enyl)-3,7-dimethylnona-2,4,6,8-tetraenamide) ( FIG. 1 ). Applicants have demonstrated that racemic VNLG-152, VNLG-153, and VNLG-147 exhibit potent antiproliferative and antimetastatic activity against androgen-sensitive and castration-resistant human prostate cancer cells in vitro. 47 Applicants have also shown that VNLG-152R (Compound 2) exerts anticancer and anti-metastatic activity in in vivo models of breast and prostate cancer and have demonstrated the pharmacokinetics of the compound, including oral bioavailability, in mice. 55~56 .

[0038] Deuterated Retinoid Compounds Hydrogen (H2O) for Altering Pharmacokinetics and Metabolism of Small Molecule Drugs 1 H, H) deuterium 2 H, D) The utility of substitution is a valuable approach in medicinal chemistry and drug discovery and development. 57~59 In fact, the first classic example was deuterobenazine, approved by the U.S. Food and Drug Administration (FDA) in 2017 for the treatment of Huntington's disease. 57、60~62 , a deuterated C currently being investigated in a Phase 3 trial for treating geographic atrophy (GA) secondary to age-related macular degeneration (AMD). 20 Over 20 deuterated drugs are currently in clinical development, including -D3-vitamin A (ALK-001). 57、60、69 .

[0039] The present disclosure provides novel deuterated analogs of Applicant's novel ATRA retinoid compounds. Applicant hypothesizes that deuterium substitution at potential metabolic hotspots of retinoid compounds, including VNLG-152R (Compound 2), can slow the in vivo metabolism of the compounds, allowing for improved pharmacokinetic properties. Indeed, modest half-life improvements (extension of residence time) of short half-life compounds can dramatically reduce effective doses. 56、64 .

[0040] To design possible deuterated analogs of VNLG-152R (compound 2), applicants identified atoms of VNLG-152R (compound 2) that are most likely to be oxidized by Cyp3A4, Cyp2D6, and Cyp2C9 as marked with circles (◯). Other predicted metabolic sites are marked with squares (□) and triangles (△), representing amide bond cleavage and aromatic ring hydroxylation to form phenols, respectively. Figure 2 maps the predicted metabolic sites onto VNLG-152R.

[0041] Applicant has discovered that amide hydrogen atoms are easily exchangeable, and based on this discovery, has focused on replacing C4-H, the three hydrogens of C4-imidazole, and the four hydrogens of amide benzyl fluoride. The possible exchanges of hydrogen with deuterium in the three regions of VNLG-152R (compound 2) translate into seven (3 factorials (3!, i.e., 3×2×1=6)+1) deuterated analogs of VNLG-152R (compound 2). Thus, the present disclosure provides a deuterated retinoid of formula A, or a pharma- ceutically acceptable salt thereof: [ka] to provide.

[0042] In formula A, R 1 ~R 8 is independently H or D, and R 1 ~R 8 At least one of R is D. 4 is D. In some embodiments, R 4 , and R 6 ~R 8 is D. In some embodiments, the deuterated retinoid is at least one selected from the following, or at least one pharma- ceutically acceptable salt selected from the following: [ka] [ka] [ka] It is.

[0043] The structures and accurate masses of deuterated compounds 3-9 are shown in FIG.

[0044] Synthesis and preparation of VNLG-152R VNLG-152R (compound 2) was previously synthesized in one step from Applicant's initial lead compound VN / 14-1 following treatment with commercially available 4-fluoroaniline in the presence of 1-hydroxybenzotriazole (HOBT) and dicyclohexylcarbodiimide (DCC) in dimethylformamide at room temperature. For clarity, the synthesis of VNLG-152R (compound 2) involving five steps from commercially available ATRA and the yield of each step is shown in scheme 1 of FIG. 4. Thus, the overall yield of the synthesis of VNLG-152R (compound 2) from ATRA is 18%. The relatively low yields of step 2 involving allylic oxidation of compound 10 with manganese dioxide and step 6 forming the DCC amide have the most significant impact on the low overall yield of VNLG-152R (compound 2).

[0045] Due to the further ongoing development of VNLG-152R (compound 2) towards clinical trials in humans, and due to the need to produce new analogs of VNLG-152R (compound 2) with enhanced pharmacokinetics, and as a strategy to generate completely novel chemical entities, Applicants have designed a significantly improved procedure for the synthesis of VNLG-152R (compound 2).

[0046] Recently, 4-oxo-ATRA (compound 15) has become commercially available. Based on previous experience with the synthesis of VNLG-152R (compound 2) from ATRA, the applicants hypothesize that the use of 4-oxo-ATRA (compound 15) as the starting material eliminates the low-yield allylic oxidation step in Scheme 1 (Figure 4). The novel synthesis of VNLG-152R (compound 2) is outlined in Scheme 2 of Figure 5.

[0047] 4-Oxo-retinamide (compound 16) is synthesized by coupling 4-oxo-ATRA (compound 15) with 4-fluoroaniline via the active ester method using 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC.HCl) and 1-hydroxybenzotriazole (HOBt). This reagent offers an advantage over the previously used DCC method because of the ease of workup without the need for column chromatographic purification to obtain the pure 4-oxo-retinamide intermediate (compound 16) in excellent yield (87%). Reduction of compound 16 with sodium borohydride (NaBH4) using a solvent mixture of methanol / tetrahydrofuran in a 2:1 ratio provides the key intermediate (±)-4-hydroxyretinamide (compound 17) in nearly quantitative (97%) yield. The production of compound 17 does not require column chromatographic purification. Finally, treatment of compound 17 with carbonyldiimidazole (CDI) using a solvent mixture of acetonitrile / tetrahydrofuran in a 1:1 ratio at room temperature affords the desired VNLG-152R (compound 2) in 56.5% yield after chromatographic purification. In this final step, applicants have developed an efficient chromatographic purification method to remove colored / unidentified impurities that have a retention time very close to that of VNLG-152R (compound 2). The novel scheme 2 synthesizes VNLG-152R (compound 2) in 48% overall yield, a substantial improvement over the 18% overall yield possible with scheme 1.

[0048] Synthesis and Preparation of Deuterated Retinoid Compounds The present disclosure also provides a method for synthesizing the deuterated analog of ATRA.The deuterated compounds disclosed in the present disclosure can be synthesized by the pathways described below and in the accompanying drawings.The materials used in the pathways described are commercially available or prepared by synthetic methods generally known in the art.These schemes are not limited to the compounds listed, nor are they limited by any particular substituents used for illustrative purposes.

[0049] Deuterated compounds 3-9 are synthesized via a three-step synthetic route as shown in Scheme 2 of the parent VNLG-152R (compound 2).

[0050] In one aspect, the disclosure provides a method for synthesizing compounds 3-5 according to the route shown in Scheme 3 of FIG. 6. The key intermediate for synthesizing the deuterated analog compounds 3-5 is 4-oxo-retinamide (compound 8). As shown in Scheme 3 of FIG. 6, 4-oxo-retinamide (compound 8) is treated with either NaBH4 or NaBD4 to give the corresponding non-deuterated 4-hydroxy compound 17 and 4H-deuterated 4-hydroxy compound 18. Treatment of compound 17 with 1,1'-carbonyldiimidazole-d6 at room temperature gives the desired (±)-4-(imidazolyl-2''3''5''-D3)-4-(H)-(4'-fluoro(phenyl))-(E)-retinamide (compound 3). Treatment of compound 18 with 1,1′-carbonyldiimidazole or 1,1′-carbonyldiimidazole-d6 affords the desired (±)4-(imidazolyl)-4-(D)-(4′-fluoro(phenyl))-(E)-retinamide (compound 4) and (±)4-(imidazolyl-2″3″5″-D3)-4-(D)-(4′-fluoro(phenyl))-(E)-retinamide (compound 5), respectively.

[0051] In another aspect, the present disclosure provides a method for synthesizing compounds 6-9 according to the route shown in Scheme 4 of FIG. 7. Deuterated analog compounds 6-9 are synthesized similarly to compounds 3-5 of Scheme 3, except that the key intermediate in the synthesis of compounds 6-9 is 4-oxo-(4'-fluoro(phenyl-2',3'5'6'-D4)-(E)-retinamide (compound 19) instead of 4-oxo-retinamide (compound 8), as shown in Scheme 4 of FIG. 7. Notably, the final step yields of all deuterated analogs were modest, which Applicant hypothesizes is due to the poor solubility of the deuterated reagents.

[0052] Applicants have developed a new and improved procedure for the synthesis of advanced VNLG-152R (compound 2). The improved synthesis and yield of VNLG-152R allows for the synthesis of novel deuterated analogs of VNLG-152R, compounds 3-9. The novel deuterated analogs of the present disclosure have been found to be potent Mnk1 degradation inducers with resulting depletion of peIF4E and other oncogenic proteins. Importantly, the in vivo pharmacokinetics of both non-deuterated VNLG-152R (compound 2) and the deuterated analogs (compounds 3-9) show that the pharmacokinetic parameters of the deuterated analogs, particularly compounds 4, 8, and 9, are significantly higher than those of the deuterated analogs, at C max , T 1 / 2 , and are superior to non-deuterated VNLG-152R in terms of AUC and MRT. The therapeutic potential of the disclosed deuterated analogs is promising. Importantly, the deuterated strategy described in this disclosure generates entirely novel chemical entities, and the new compositions will undoubtedly support further development of these promising deuterated compounds.

[0053] Pharmaceutical Compositions The deuterated retinoids or salts thereof of the present disclosure can be used to prepare pharmaceutical compositions effective for treating conditions associated with Mnk1 / 2 signaling and / or androgen signaling, including, for example, breast cancer, prostate cancer, bladder cancer, pancreatic cancer, hepatocellular carcinoma, benign prostatic hyperplasia (BPH), and Kennedy's disease (spinal-bulbar muscular atrophy), and blood cancers. In some embodiments, the deuterated compounds or salts thereof of the present disclosure are used to prepare pharmaceutical compositions effective for treating triple-negative breast cancer.

[0054] The present disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the above-mentioned deuterated retinoid compounds.The deuterated retinoid compounds (also referred to as "active compounds" in the present disclosure) or salts thereof of the present disclosure can be incorporated into pharmaceutical compositions suitable for administration.Such compositions typically comprise a deuterated retinoid compound or salts thereof and a pharma- ceutically acceptable carrier.

[0055] The pharmaceutical composition comprises a deuterated retinoid compound of formula A, or a pharma- ceutically acceptable salt thereof: [ka] may include.

[0056] In formula A, R 1 ~R 8 is independently H or D, and R 1 ~R 8 At least one of R is D. 4 is D. In some embodiments, R 4 , and R 6 ~R 8 At least one of is D.

[0057] In some embodiments, the pharmaceutical composition comprises at least one of compounds 3-9 or a salt thereof. In some embodiments, the pharmaceutical composition comprises at least one of compounds 4, 8, and 9 or a salt thereof. In some embodiments, the pharmaceutical composition comprises a mixture of at least one ATRA retinoid compound or a salt thereof and at least one of compounds 3-8 or a salt thereof.

[0058] As used in this application, "pharmaceutically acceptable carrier" refers to a component in a particular dosage form that is considered inert and is typically used in the pharmaceutical field to formulate a dosage form containing a particular active compound. Pharmaceutically acceptable carriers include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, etc., that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutical active substances is well known in the art. Any conventional media or agent is contemplated for use in the composition, except insofar as it is incompatible with the active compound. Auxiliary active compounds can also be incorporated into the composition.

[0059] The deuterated retinoid compounds of the present disclosure can be used as a monotherapy or in combination with other therapeutic agents. The deuterated retinoid compounds or salts thereof can be used in combination with other cancer treatments and drugs. For example, the deuterated retinoid compounds or salts thereof of the present disclosure can be used as part of or in combination with known cancer treatments, such as hormone therapy, chemotherapy, radiation therapy, immunotherapy, and / or surgery. The deuterated retinoid compounds or salts thereof of the present disclosure can also be used in combination with one or more known available drugs or other compounds. The therapeutic efficacy of one of the deuterated retinoid compounds or salts thereof described in the present disclosure can be enhanced by the administration of an adjuvant (i.e., the adjuvant itself may have minimal therapeutic benefit, but when combined with another therapeutic agent, the overall therapeutic benefit to the patient is enhanced). Or, by way of example only, the benefit experienced by the patient can be increased by the administration of one of the deuterated retinoid compounds or salts thereof of the present disclosure together with another therapeutic agent (including a treatment regimen) that also has a therapeutic benefit. In either case, regardless of the disease or condition being treated, the overall benefit experienced by the patient may be the synergistic effect of the multiple therapeutic agents, or the patient may experience a synergistic benefit. When the deuterated retinoid compounds or salts thereof of the present disclosure are administered in conjunction with other therapeutic agents, the dosage of the co-administered therapeutic agent will of course vary depending on the type of co-drug used, the specific drug used, the disease or condition being treated, and the like. Furthermore, when co-administered with one or more biologically active agents, the deuterated retinoid compounds or salts thereof of the present disclosure may be administered simultaneously with the biologically active agent, or sequentially. When administered simultaneously, the multiple therapeutic agents may be provided in a single, unified form, or in multiple forms (by way of example only, as a single pill, or as two separate pills). One of the therapeutic agents may be given as multiple doses, or both may be given as multiple doses. When administered sequentially, the attending physician will determine the appropriate regimen. Furthermore, the combination methods, compositions, and formulations should not be limited to the use of only two agents. Multiple therapeutic combinations are envisioned.

[0060] As used in this application, a "therapeutically effective amount" of a deuterated retinoid compound or a salt thereof is an amount that effectively achieves a desired therapeutic result in a subject. Such an amount can be initially determined by knowledge in the art, by performing in vitro tests, by performing metabolic tests in healthy laboratory animals, and / or by performing clinical trials. Of course, the dosage of the various deuterated retinoid compounds or their salts of the present disclosure will vary somewhat depending on, among other things, the host being treated, the particular mode of administration. Those skilled in the art can determine the optimal dosage of the deuterated retinoid compound or its salt of the present disclosure selected based on clinical experience and treatment indication.

[0061] The pharmaceutical composition according to the present disclosure is configured to facilitate administration of a deuterated retinoid compound or a salt thereof to a subject. The deuterated retinoid compound or a salt thereof of the present disclosure, or a pharmaceutical composition containing a deuterated retinoid compound or a salt thereof, can be administered according to any suitable method or route, including, but not limited to, intranasal, intramuscular, intratracheal, subcutaneous, intradermal, transdermal, sublingual, topical application, intravenous, ocular (e.g., topical to the eye, intravitreal, etc.), rectal, nasal, oral, topical administration, and other enteral and parenteral administration routes.

[0062] In some embodiments, the pharmaceutical composition is formulated for oral administration. In some embodiments, the pharmaceutical composition may contain a sufficient amount of the deuterated retinoid compound or a salt thereof to administer 0.01-100 mg / kg body weight per day, e.g., 0.01-35 mg / kg body weight per day, 0.05-20 mg / kg body weight per day, or 5-20 mg / kg body weight per day of the deuterated retinoid compound or a salt thereof.

[0063] Compositions suitable for oral administration can contain (a) liquid solutions, such as an effective amount of one or more of the disclosed compounds or salts dissolved in a diluent, such as water or saline; (b) capsules, sachets or tablets, each containing a predetermined amount of the active ingredient as a solid or granules; (c) suspensions in a suitable liquid; and (d) suitable emulsions. For oral administration, the disclosed deuterated retinoids or salts thereof can be formulated, for example, by combining the active compound with pharma- ceutically acceptable carriers or excipients known in the art. Such carriers allow the disclosed retinamides to be formulated as tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, and the like, for oral ingestion by the patient to be treated.

[0064] In some embodiments, pharmaceutical compositions are formulated for parenteral administration.Compositions suitable for parenteral administration include aqueous and non-aqueous solutions that can contain antioxidants, buffers, bacteriostatic agents, and solutes that make the formulation isotonic with the blood of intended recipients, isotonic sterile injection solutions, and aqueous and non-aqueous sterile suspensions that can contain suspending agents, solubilizers, thickening agents, stabilizers, and preservatives.The preparations can be provided in unit-dose or multi-dose sealed containers such as ampoules and vials, and can be stored in a lyophilized state that only requires the addition of sterile liquid carriers for injection, such as water, immediately before use.Extemporaneous injection solutions and suspensions can be prepared from the above-mentioned types of sterile powders, granules, and tablets.

[0065] Delivery of the deuterated retinoid compound or salt thereof can be enhanced by facilitating a more pharmacologically effective amount of the compound to reach the site of action. Delivery can also be enhanced by facilitating more efficient delivery of the compound across cell membranes or across intracellular and intracellular spaces.

[0066] Non-limiting examples of suitable carriers or excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose.The formulation can further include lubricants such as talc, magnesium stearate, and mineral oil, wetting agents, emulsifying and suspending agents, preservatives such as methyl hydroxybenzoate and propyl hydroxybenzoate, sweeteners, and flavoring agents.

[0067] The deuterated retinoid compound or its salt of the present disclosure can be applied to exert local or systemic effects, or both.For example, when systemic delivery is desired, administration can involve invasive administration of pharmaceutical preparations or topical or mucosal administration that is systemically absorbed.The pharmaceutical composition containing the deuterated retinoid compound or its salt of the present disclosure can be administered in a targeted drug delivery system.Furthermore, the pharmaceutical composition containing the deuterated retinoid compound or its salt of the present disclosure can be formulated to be a rapid release formulation, a sustained release formulation, or an intermediate release formulation according to suitable procedures known in the art.

[0068] The present disclosure provides a method for preparing a pharmaceutical composition containing the deuterated retinoid compound or its salt of the present disclosure. Such a composition can further comprise an additional active agent. Thus, the present disclosure further provides a method for preparing a pharmaceutical composition by formulating a pharma- ceutically acceptable carrier with the deuterated retinoid compound of the present invention or its salt and one or more additional active compounds.

[0069] Treatment The present disclosure also provides methods of treating various conditions, particularly conditions associated with Mnk1 / 2 and / or androgen signaling. In some embodiments, the methods involve administering a therapeutically effective amount of a deuterated retinoid compound or salt thereof of the present disclosure, or a pharmaceutical composition comprising a therapeutically effective amount of a deuterated retinoid compound or salt thereof, to a subject (e.g., a human or non-human mammal) in need thereof. In some embodiments, the deuterated retinoid compound or salt thereof of the present disclosure is administered to treat at least one selected from the group consisting of breast cancer, prostate cancer, bladder cancer, pancreatic cancer, hepatocellular carcinoma, benign prostatic hyperplasia (BPH), and Kennedy's disease (spinal-bulbar muscular atrophy), and blood cancer. In some embodiments, the condition is triple-negative breast cancer. The deuterated retinoid compound may include at least one of compounds 3-9, or salts thereof. In some embodiments, the deuterated retinoid may include at least one of compounds 4, 8, and 9, or salts thereof.

[0070] In some embodiments, the deuterated retinoid compound or salt thereof is administered orally or parenterally.

[0071] A "therapeutically effective amount" is as defined above. In the context of the present invention, the dose administered to an animal, particularly a human, should be sufficient to affect a therapeutic response in the animal or human over a reasonable time frame. The specific dose level and frequency of administration may vary depending on a variety of factors, including the activity of the specific active compound, its metabolic stability and length of action, excretion rate, mode and time of administration, the subject's age, weight, health, sex, diet, etc., as well as the severity of, for example, breast cancer. Any effective amount of the compound may be administered, for example, about 1 mg to about 500 mg per day, about 50 mg to about 150 mg per day, etc. In some embodiments, the deuterated retinoid compound or salt thereof may be administered at 0.01 to 100 mg / kg body weight per day, for example, 0.01 to 35 mg / kg body weight per day, 0.05 to 20 mg / kg body weight per day, or 5 to 20 mg / kg body weight per day of the deuterated retinoid compound or salt thereof. The deuterated retinoid compounds or salts of the present disclosure can be administered in any form and in such dosages by any effective route, including, for example, oral, parenteral, enteral, intraperitoneal, topical, transdermal (e.g., using any standard patch), ophthalmic, nasal, topical, parenteral, e.g., aerosol, spray, inhalation, subcutaneous, intravenous, intramuscular, buccal, sublingual, rectal, vaginal, intraarterial, and intrathecal, etc. In some embodiments, the deuterated retinoid compound or salt thereof is administered once daily.

[0072] The deuterated retinoid compound or salt of the present disclosure may be used in combination with procedures that may provide additive or synergistic benefits to patients.The deuterated retinoid compound or salt of the present disclosure, whether as a monotherapy or in combination therapy, may be administered before, during or after the onset of the disease or condition to be treated, and the timing of administering the composition containing the deuterated retinoid or salt may vary.Thus, the deuterated retinoid compound or salt may be administered to the subject during or as soon as possible after the onset of symptoms.The deuterated retinoid compound or salt is preferably administered as soon as practicable after the onset of the disease or condition listed in the present disclosure is detected or suspected, for the period required for the treatment of the disease.The length of treatment may vary from subject to subject, and the length may be determined using known criteria. Administration can be acute (e.g., short term (e.g., a single dose, administration for 1 day to 1 week)) or chronic (e.g., long term (e.g., administration for more than 1 week, from about 2 weeks to about 1 month, from about 1 month to about 3 months, from about 3 months to about 6 months, from about 6 months to about 1 year, or for more than 1 year)).

[0073] As used in this application, "treat" and all its forms and tenses (including, for example, treat, treating, treated, and treatment) refers to therapeutic and / or prophylactic or preventative treatment. Those in need of treatment include those already having a pathological condition enumerated in this disclosure and those in which a pathological condition is to be prevented. For example, "treat" means to medically or surgically alter, apply, bring about, ameliorate, take care of, or deal with, improve, cure, stop, and / or prevent an undesirable biological (pathogenic) process. One of skill in the art will recognize that treatment may or may not be curative. EXAMPLES

[0074] The following examples are provided to illustrate the present invention and to assist one of ordinary skill in the relevant art in making and using the present invention, and are not intended to limit the scope of the invention.

[0075] Example 1 All materials listed below were purchased and used without further purification. Starting material 4-oxo-ATRA was purchased from Synnovator, Inc., 11 Centrewest Ct, Cary, NC 27513, USA. 1,1'-carbonyldiimidazole-d6 and 4-fluoroaniline-2,3,5,6-d4 were purchased from CDN Isotopes Inc., 88 Leacock, Poine-Claire, QC H9R 1H1, Canada. All other reagents and solvents of highest purity research or spectrophotometric grade were commercially available from Sigma-Aldrich. Silica gel plates (Merck F254) were used for thin layer chromatography (TLC) and developed with a mixture of ethyl acetate (EtOAc) / petroleum ether (1:3) or 1% ethanol in EtOAc unless otherwise specified, and visualized with light at 254 nm and 365 nm. Petroleum ether refers to a light oil with a boiling point (bp) of 40-60 °C. Column chromatographic purification was performed on a CombiFlash Teledyne ISCO automated chromatography instrument using normal phase silica columns. 1 H and 13 C NMR spectra were recorded in CDCl3 or DMSO-d6 using a Bruker 400 MHz NMR instrument and chemical shifts are reported in ppm on the δ scale relative to tetramethylsilane. 2H NMR spectra were recorded in 0.5% DMSO-d6 in DMSO using a Bruker Avance III 950 MHz NMR instrument. High-resolution mass spectra were obtained on a Bruker 12T APEX-Qe FTICR-MS instrument in positive ion ESI mode by Cosmic Laboratory, Old Dominion University, Norfolk, VA. Melting points (mp) were measured on a Fischer Johns melting point apparatus uncorrected. Compound purity was determined by reversed phase on a Waters Acquity UPLC LC system equipped with a photodiode array detector using a Novapac-C18 (3.9 × 150 mm × 4 μm) column as the stationary phase. For the mobile phase, solvent A contained 20 mM ammonium acetate in methanol-water (70:30 v / v), solvent D was methanol, the flow rate was 0.5 mL / min, and a gradient flow was maintained (see chromatograms in the supplementary section for gradient flow details). The column was kept at room temperature and the injection volume was 10.0 L. The purity of all final compounds was determined to be at least 95% pure by a combination of UPLC, NMR and HRMS.

[0076] General Method A: Synthesis of 4-oxo-retinamides (compounds 16 and 19) from 4-oxo-ATAR (compound 15) In general method A, to a suspension of 4-oxo-ATRA (compound 15) (1 molar ratio (MR)), 1-hydroxybenzotriazole (2 MR), 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC.HCl) (2 MR), dimethylformamide (5 mL / g of substrate) is added N,N-diisopropylethylamine (10 MR) and 4-fluoroaniline or 4-fluoroaniline-2,3,5,6-d4 (1.1 MR). The reaction mixture is sonicated for 5 minutes to obtain a homogeneous solution and then stirred at room temperature for 18 hours. Excess diisopropylethylamine from the reaction mixture is removed under vacuum. To the remaining mixture, approximately 25 mL of ice / water is added and stirred vigorously until a fine solid is obtained, then an additional 25 mL of water is added (10 mL / mL of DMF). The product is filtered at room temperature, washed thoroughly with water, filtered and dried. The resulting orange solid is recrystallized from hot ethanol (2 mL / g of product) and filtered at room temperature to give orange needles which are used in the next step of the synthetic scheme without further purification.

[0077] Example 2 General Method B: Conversion of 4-oxo-retinamides (compounds 16, 19) to hydroxy-retinamides (compounds 17, 18, 20, and 21) Reagents, materials, and equipment are as described in Example 1. In general method B, a solution of 4-oxo-retinamide (compounds 16, 19) (1 MR), methanol (10 mL / g of substrate), and tetrahydrofuran (5 mL / g of substrate) is cooled to ice temperature (about 4° C.). Sodium borohydride or sodium borodeuteride (1.2 MR) is added in three portions over 15 minutes, and the reaction mixture is stirred at room temperature for 30 minutes. To the resulting yellow solution (orange-yellow), acetone (2 mL / g of substrate) is added and stirred for 5 minutes. The reaction mixture is then concentrated under vacuum, treated with ice water, filtered, and dried under vacuum. This product is used in the next step of the synthetic scheme without further purification.

[0078] Example 3 General Method C: Addition of Imidazole at C-4 Position (1, 10-16) to Synthesize VNLG-152R (Compound 2) Reagents, materials, and equipment are as described in Example 1. The following general method C is used to synthesize VNLG-152R (compound 2) from intermediate 4-hydroxy-retinamides (compounds 17, 18, 20, 21) by adding imidazole at the C-4 position. In general method C, 1,1'-carbonylimidazole or 1,1'-carbonyldiimidazole-d6 (1.5 MR) is added to a solution of 4-hydroxy-retinamide (compounds 17, 18, 20, 21) (1 MR) in acetonitrile (5 mL / g of substrate) and tetrahydrofuran (5 mL / g of substrate) and stirred at room temperature for 1.5 hours. The initial yellow solution becomes orange. The solvent is evaporated under vacuum, reconstituted with dichloromethane (3 mL / g of substrate), water is added (6 ml / g of substrate), and the dichloromethane is removed from the mixture under vacuum. This process is repeated twice to solubilize the unreacted CDI and imidazole in the aqueous phase. Filter the resulting solid or decant the water if the residue is sticky and dry the contents under vacuum. Purify the crude product on a CombiFlash automated chromatography instrument by a silica gel column (5 inches) using solvent A (ethyl acetate) for 2 min, then solvent B (95:5:0.5 ethyl acetate / ethanol / trimethylamine) at 0-5% over 20 min. Combine the fractions between 10.5 and 18 min and evaporate to give a yellow solid.

[0079] Example 4 (±) 4-Imidazolyl-4(H)-(4'-fluoro(phenyl))-(E)-retinamide (VNLG-152R, compound 2) Compound 2 (VNLG-125R) was synthesized using general method C from Example 3. To a solution of (±)4-hydroxy-4-[H]-(4'-fluoro(phenyl)-(E)-retinamide (compound 17) (0.94 g, 2.29 mmol) in acetonitrile (5 mL) and tetrahydrofuran (5 mL), 1,1'-carbonyldiimidazole (0.56 g, 3.44 mmol) was added and stirred at room temperature for 1.5 h (the initial yellow solution turns orange). The solvent was evaporated under vacuum, reconstituted with dichloromethane (3 mL), water was added, and the mixture was stripped of dichloromethane under vacuum. This process was repeated twice. The mixture was washed twice to solubilize the unreacted CDI and imidazole in the aqueous phase. The resulting solid was filtered (0.9 g) and purified on a CombiFlash automated chromatography instrument using a silica gel column (5 inches) with solvent A (ethyl acetate) for 2 min, followed by solvent B (95:5:0.5 ethyl acetate / ethanol / trimethylamine) at 0-5% over 20 min. The fractions between 10.5-18 min were combined and evaporated to give a yellow solid. 0.52 g (49%), mp, calcined at 105°C, melted at 111-112°C, 1 H NMR (400 MHz, CDCl3) δ ppm 1.03 - 1.19 (m, 6 H) 1.45 - 1.59 (m, 2 H) 1.59 - 1.64 (m, 3 H) 1.81 - 1.89 (m, 1 H) 2.02 (s, 3 H) 2.07 - 2.16 (m, 1 H) 2.31 - 2.53 (m, 3 H) 4.54 (t, 1 H) 5.88 (br. s., 1 H) 6.06 - 6.22 (m, 2 H) 6.22 - 6.37 (m, 2 H) 6.93 (s, 1 H) 6.93 - 7.06 (m, 3 H) 7.09 (s, 1H) 7.52 (s, 1 H) 7.55 (br. s., 2 H) 8.14 (br. s., 1 H); 1H NMR (400 MHz, DMSO-d6) δ ppm 1.08 (s, 3H) 1.11 (s, 3 H) 1.40 - 1.59 (m, 5 H) 1.73 - 1.82 (m, 1 H) 2.01 (s, 3 H) 2.02 - 2.12 (m, 1 H) 2.23 - 2.41 (m, 3 H) 4.72 (t, 1 H) 6.02 (s, 1 H) 6.28 (m, 2 H) 6.31 - 6.47 (m, 2 H) 6.92 (s, 1 H) 7.01 (dd, 1 H) 7.07 - 7.20 (m, 3 H) 7.62 (s, 1 H) 7.66 (dd, 2 H) 10.07 (s, 1 H). 13 C NMR (101 MHz, DMSO-d6) δ ppm 13.0, 13.7, 18.8, 28.1, 28.3, 29.1, 34.7, 35.1, 57.4, 115.5, 115.8, 118.8, 121.0, 121.1, 123.2, HRMS Calculated value 460.2758 (C 29 H 34 FN3O H+) measured value 460.2760.

[0080] Example 5 4-Oxo-(4'-fluoro(phenyl))-(E)-retinamide (Compound 16) Compound 16 was synthesized using general method A of Example 1. To a suspension of 4-oxo-ATRA (compound 15) (2 g, 6.36 mmol), 1-hydroxybenzotriazole (1.71 g, 12.7 mmol), 1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride (EDC.HCl) (2.44 g, 12.7 mmol), dimethylformamide (10 mL) was added N,N-diisopropylethylamine (8.22 g, 63.6 mmol) and 4-fluoroaniline (0.78 g, 7.0 mmol) and sonicated for 5 minutes to obtain a homogeneous solution. The mixture was then stirred at room temperature for 18 hours. Excess diisopropylethylamine was removed under vacuum. To the remaining mixture, about 50 mL of ice / water was added and stirred vigorously until a fine solid was obtained, then another 50 mL of water was added. The product was filtered at room temperature, washed thoroughly with water, filtered and dried. The resulting orange solid (2.49 g) of 4-oxo-(4'-fluoro(phenyl)-(E)-retinamide (compound 16) was recrystallized using hot ethanol (5 mL) to give orange needles. 2.26 g (87%), mp, sintered at 166 °C, melted at 171 °C. 1 H NMR (400 MHz, CDCl3) δ ppm 1.20 (s, 6 H) 1.79 - 1.92 (m, 5 H) 2.04 (s, 3 H) 2.43 (s, 3 H) 2.52 (t, 2 H) 5.87 (s, 1 H) 6.26 (d, 1 H) 6.30 - 6.41 (m, 3 H) 6.92 - 7.07 (m, 3 H) 7.40 (br. s., 1 H) 7.54 (br. s., 2 H); 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.17 (s, 6 H) 1.78 (s, 3 H) 1.79 - 1.93 (m, 2 H) 2.04 (s, 3 H) 2.36 (s, 3 H) 2.43 (t, 2 H) 6.06 (s, 1 H) 6.33 - 6.55 (m, 4 H) 7.03 (dd, 1 H) 7.14 (t, 2 H) 7.59 - 7.75 (m, 2 H) 10.10 (s, 1 H). 13C NMR (101 MHz, DMSO-d6) δ ppm 12.9, 13.7, 13.9, 27.7, 34.2, 35.7, 37.3, 115.5, 115.8, 121.1, 121.1, 123.7, 126.1, 129.2, 130.0, 133.6, 136.3, 138.0, 138.0, 140.8, 148.7, 157.1, 159.5, 160.9, 165.0, 198.2.

[0081] Example 6 (±) 4-Hydroxy-4-[H]-(4'-fluoro(phenyl)-(E)-retinamide (Compound 17) Compound 17 was synthesized using general method B of Example 2. To an ice-cold orange suspension of 4-oxo-(4'-fluoro(phenyl)-(E)-retinamide (compound 16) (1 g, 2.45 mmol), methanol (10 mL) and tetrahydrofuran (5 mL) was added fresh sodium borohydride (0.123 g, 2.94 mmol) in three portions over a period of 15 min. The mixture was then stirred at room temperature for 30 min. To the resulting yellow solution was added acetone (2 mL) and stirred for 5 min. The reaction mixture was then concentrated under vacuum, treated with ice water, filtered and dried under vacuum to give a yellow solid. 0.98 g (97%), mp, 161-163 °C, 1 H NMR (400 MHz, CDCl3) δ ppm 1.05 (s, 3 H) 1.02 (s, 3 H) 1.44 (ddd, 1 H) 1.54 (d, 1 H) 1.62 - 1.76 (m, 2 H) 1.84 (s, 3 H) 1.86 - 1.97 (m, 1 H) 2.01 (s, 3 H) 2.42 (s, 3 H) 4.02 (d, 1 H) 5.80 (s, 1 H) 6.09 - 6.19 (m, 2 H) 6.19 - 6.26 (m, 1 H) 6.29 (d, 1 H) 6.90 - 7.06 (m, 3 H) 7.20 (br 1 H) 7.51 (br. s., 2 H); 1H NMR (400 MHz, DMSO-d6) δ ppm 0.90 - 1.16 (m, 6 H) 1.26 - 1.39 (m, 1 H) 1.50 - 1.69 (m, 2 H) 1.75 (s, 4 H) 1.99 (s, 3 H) 2.35 (s, 3 H) 3.82 (br. s., 1 H) 4.67 (br. s., 1 H) 6.02 (s, 1 H) 6.11 - 6.27 (m, 2 H) 6.31 (d, 1 H) 6.39 (d, 1 H) 7.01 (dd1 H) 7.14 (t, 2 H) 7.66 (dd, 2 H) 10.07 (s, 1 H). 13 C NMR (101 MHz, DMSO-d6) δ ppm 13.0, 13.7, 18.9, 28.3, 28.9, 29.2, 34.7, 35.2, 68.7, 115.5, 115.7, 121.1, 121.1, 122.9, 127.8, 130.3, 130.9, 132.3, 136.3, 136.6, 137.9, 138.6, 139.5, 148.9, 157.1, 159.5, 165.0.

[0082] Example 7 (±)4-(imidazolyl-2''3''5''-D 3 )-4-(H)-(4'-fluoro(phenyl))-(E)-retinamide (compound 3) Compound 3 was synthesized using (±)4-hydroxy-4-[H]-(4'-fluoro(phenyl)-(E)-retinamide (compound 17) (0.73 g, 1.78 mmol) and 1,1'-carbonyldiimidazole-d6 (0.45 g, 2.67 mmol) by following general method C. Yellow solid 0.37 g (44%), mp, sintered at 103 °C, melted at 115-117 °C, 11H NMR (400 MHz, DMSO-d6) δ ppm 1.11 (s, 3 H) 1.08 (s, 3 H) 1.42 - 1.55 (m, 5 H) 1.71 - 1.85 (m, 1 H) 2.01 (s, 3 H) 2.02 - 2.12 (m, 1 H) 2.29 - 2.40 (m, 3 H) 4.74 (t, 1 H) 6.03 (s, 1 H) 6.24 - 6.32 (m, 2 H) 6.32 - 6.45 (m, 2 H) 7.01 (dd, 1 H) 7.08 - 7.19 (m, 2 H) 7.60 - 7.71 (m, 2 H) 10.10 (s, 1 H); 2 1H NMR (146 MHz, 0.5% DMSO-d6, in DMSO) δ ppm 6.97 (br s, 1D) 7.13 (br s, 1D) 7.71 (br s, 1D); 13 13C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.8 , 28.0 , 28.2 , 29.1 , 34.7 , 35.0 , 39.3 , 39.5 , 39.7 , 39.9 , 40.1 , 40.4 , 40.6 , 57.6 , 115.5 , 115.7 , 121.0 , 121.1 , 123.2 , 125.6 , 126.8 , 130.2 , 131.6 , 136.3 , 136.3 , 137.0 , 138.2 , 139.1 , 144.1 , 148.8, 157.1 , 159.5 , 165.0; HRMS calculated 463.2946 (C 29 H 31 D3FN3O H+) Found 463.2949.

[0083] Example 8 (±) 4-(imidazolyl)-4-(D)-(4'-fluoro(phenyl))-(E)-retinamide (compound 4) Compound 4 was synthesized using (±)4-hydroxy-4-[D]-(4'-fluoro(phenyl)-(E)-retinamide (compound 18) (0.5 g, 1.29 mmol) and 1,1'-carbonyldiimidazole (0.31 g, 1.93 mmol) by following general method C. Yellow solid 0.27 g (47%), mp, 165-167 °C. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.09 (s, 3 H) 1.12 (s, 3 H) 1.51 (s, 4 H) 1.73 - 1.82 (m, 1 H) 2.02 (s, 3 H) 2.03 - 2.09 (m, 1 H) 2.36 (s, 3 H) 6.03 (s, 1 H) 6.25 - 6.45 (m, 4 H) 6.93 (s, 1 H) 7.02 (dd, 1 H) 7.08 - 7.18 (m, 3 H) 7.63 (s, 1 H) 7.67 (dd, 2 H) 10.08 (s, 1 H); 2 H NMR (146 MHz,0.5% DMSO-d6, in DMSO) δ ppm 4.69 (br s, 1D); 13 C NMR (101 MHz, DMSO-d6) δ ppm 12.4 , 13.1 , 18.2 , 27.4 , 27.7 , 28.6 , 34.1 , 34.5 , 115.0 , 115.2 , 118.2 , 120.5 , 120.5 ,122.6 , 125.2 , 126.3 , 128.2 , 129.6 , 131.0 , 135.7 , 135.8 , 136.4 , 136.7 , 137.7 , 138.5 , 143.4 , 148.3 , 156.5 , 158.9 , 164.4; HRMS calculated 461.2821 (C 29 H 33 DFN3O H+) Found 461.2820.

[0084] Example 9 (±)4-(imidazolyl-2''3''5''-D 3 )-4-(D)-(4'-fluoro(phenyl))-(E)-retinamide (compound 5) Compound 5 was synthesized using (±)4-hydroxy-4-[D]-(4'-fluoro(phenyl)-(E)-retinamide (compound 18) (0.5 g, 1.22 mmol) and 1,1'-carbonyldiimidazole-d6 (0.31 g, 1.82 mmol) by following general method C. Yellow solid 0.27 g (47%), mp, sintered at 125 °C, melted at 131 °C, 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.09 (s, 3 H) 1.13 (s, 3 H) 1.45 - 1.51 (m, 2 H) 1.53 (s, 3 H) 1.80 - 1.87 (m, 1 H) 2.02 (s, 3 H) 2.36 (s, 3 H) 6.09 (s, 1 H) 6.19 - 6.32 (m, 2 H) 6.32 - 6.46 (m, 2 H) 7.01 (dd, 1 H) 7.07 - 7.26 (m, 2 H) 7.56 - 7.82 (m, 2 H) 10.24 (s, 1 H); 2H NMR (146 MHz,0.5% DMSO-d6, in DMSO) δ ppm 4.90 (br s, 1D) 6.95-7.55 (br s, 2D) 7.61 (br s, 1D). 13 C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.7 , 27.3 , 28.1 , 29.0 , 34.5 , 34.8 , 115.5 , 115.7 , 121.0 , 121.1 , 123.4 , 123.8 , 126.5 , 130.1 , 131.8 , 136.4 , 137.2 , 138.1 , 139.3 , 145.8 , 148.7 , 157.1 , 159.4 , 165.0; HRMS calculated 464.3009 (C 29 H 30 D4FN3O H+) Found 434.3016.

[0085] Example 10 (±) 4-Imidazolyl-4(H)-(4'-fluoro(phenyl-2'3'5'6'-D 4 ))-(E)-Retinamide (Compound 6) Compound 6 was synthesized using (±)4-hydroxy-4-[H]-(4'-fluoro(phenyl-2'3'5'6'-D4)-(E)-retinamide (compound 20) (0.5 g, 1.21 mmol) and 1,1'-carbonyldiimidazole (0.29 g, 1.81 mmol) by following general method C to give 0.29 g (51%) of a yellow solid, sintered at 105 °C, melted at 113-115 °C, 11H NMR (400 MHz, DMSO-d6) δ ppm 1.08 (s, 3 H) 1.11 (s, 3 H) 1.45 - 1.51 (m, 5 H) 1.74 - 1.82 (m, 1 H) 2.01 (s, 3 H) 2.05 (d, 1 H) 2.35 (s, 3 H) 4.74 (br. s., 1 H) 6.02 (s, 1 H) 6.28 (m, 2 H) 6.33 - 6.43 (m, 2 H) 6.97 (s, 1 H) 6.99 - 7.06 (m, 1 H) 7.14 (s, 1 H) 7.72 (br. s., 1 H) 10.08 (s, 1 H); 2 1H NMR (146 MHz, 0.5% DMSO-d6, in DMSO) δ ppm 7.15 (br s, 2D) 7.67 (br s, 2D); 13 13C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.8 , 28.0 , 28.2 , 29.1, 34.7 , 35.0 , 57.6 , 118.9 , 123.2 , 125.6 , 126.8 , 128.2 , 130.2 , 131.6 , 136.2 , 137.0 , 137.2 , 138.2 , 139.1 , 144.1 , 148.8 , 157.0 , 159.4 , 165.0; 464.3009 (C 29 H 30 D4FN3O H+) Measured value 464.3010.

[0086] Example 11 (±)4-(imidazolyl-2''3''5''-D3 )-4-(H)-(4'-fluoro(phenyl-2'3'5'6'-D 4 ))-(E)-Retinamide (Compound 7) By following the general method C, compound 7 was synthesized using (±)4-hydroxy-4-[H]-(4'-fluoro(phenyl-2'3'5'6'-D4)-(E)-retinamide (compound 20) (0.5 g, 1.21 mmol) and 1,1'-carbonyldiimidazole-d6 (0.31 g, 1.81 mmol) to give 0.15 g (27%) of a yellow solid. mp, sintered at 105 °C, melted at 113-115 °C, 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.08 (s, 3 H) 1.11 (s, 3 H) 1.47 (d, 2 H) 1.48 - 1.51 (m, 3 H) 1.77 (dd, 1 H) 2.01 (s, 3 H) 2.06 (d, 1 H) 2.35 (s, 3 H) 4.73 (t, 1 H) 6.03 (s, 1 H) 6.27 - 6.45 (m, 4 H) 7.01 (dd, 1 H) 10.10 (s, 1 H); 2 H NMR (146 MHz,1% DMSO-d6, in DMSO) δ ppm 6.91 (br s, 1D) 6.99-7.31 (br s, 3D) 7.5-7.89 (br s, 3D). 13 C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.4 , 18.7 , 18.8 , 27.9 , 28.1 , 28.3 , 29.1 , 34.6 , 34.7 , 35.1 , 57.4 , 123.2 , 125.8 , 126.9 , 130.2 , 131.6 , 136.1 , 137.0 , 138.2 , 139.0, 143.9 , 148.8 , 157.0 , 159.4, 165.0; HRMS calculated 467.3198 (C 29 H 27 D7FN3O H+) Found 467.30198.

[0087] Example 12 (±) 4-(imidazolyl)-4-(D)-(4'-fluoro(phenyl-2'3'5'6'-D 4 ))-(E)-Retinamide (Compound 8) Compound 8 was synthesized using (±)4-hydroxy-4-[D]-(4'-fluoro(phenyl-2'3'5'6'-D4)-(E)-retinamide (compound 21) (0.5 g, 1.21 mmol) and 1,1'-carbonyldiimidazole (0.29 g, 1.81 mmol) by following general method C to give 0.24 g (42%) of a yellow solid. mp, sintered at 95 °C, melted at 112 °C, 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.08 (s, 3 H) 1.11 (s, 3 H) 1.45 - 1.52 (m, 5 H) 1.74 - 1.80 (m, 1 H) 2.01 (s, 3 H) 2.03 - 2.08 (m, 1 H) 2.35 (s, 3 H) 6.03 (s, 1 H) 6.20 - 6.47 (m, 4 H) 6.90 - 6.95 (m, 1 H) 7.01 (dd, 1 H) 7.10 (m, 1 H) 7.62 (s, 1 H) 10.10 (s, 1 H); 2 H NMR (146 MHz,0.5% DMSO-d6, in DMSO) δ ppm 4.69 (br s, 1D) 7.15 (br s, 2D) 7.67 (br s, 2D). 13 C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.8 , 27.9 , 28.1 , 28.2 , 29.1 , 34.7 , 35.0, 118.7 , 123.2 , 125.7 , 125.8 , 126.9, 128.7 , 130.2 , 131.6 , 136.2 , 137.0 , 137.3 , 138.2 , 139.0 , 144.0 , 144.0 , 148.8 , 157.0 , 159.4 , 165.0; HRMS calculated 465.3072 (C 29 H 29 D5FN3O H+) Found 465.3075.

[0088] Example 13 (±)4-(imidazolyl-2''3''5''-D 3 )-4-(D)-(4'-fluoro(phenyl-2'3'5'6'-D 4 ))-(E)-Retinamide (Compound 9) Compound 9 was synthesized using (±)4-hydroxy-4-[D]-(4'-fluoro(phenyl-2'3'5'6'-D4)-(E)-retinamide (compound 21) (0.5 g, 1.21 mmol) and 1,1'-carbonyldiimidazole-d6 (0.3 g, 1.81 mmol) by following general method C to give 0.135 g (24%) of a yellow solid. mp, sintered at 95 °C, melted at 108 °C, 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.08 (s, 3 H) 1.11 (s, 3 H) 1.46 - 1.49 (m, 2 H) 1.50 (s, 3 H) 1.74 - 1.80 (m, 1 H) 2.01 (s, 3 H) 2.03 - 2.08 (m, 1 H) 2.32 - 2.37 (s, 3 H) 6.03 (s, 1 H) 6.28 (s, 2 H) 6.33 - 6.44 (m, 2 H) 7.01 (dd, 1 H) 10.05 - 10.12 (m, 1 H); 2H NMR (146 MHz,0.5% DMSO-d6, in DMSO) δ ppm 4.70 (br s, 1D) 6.94 (br s, 1D) 7.10-7.32 (br s, 3D) 7.40-7.92 (br s, 3D). 13 C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.8 27.98 , 28.1 , 28.2 , 29.1 , 34.7 , 35.1 , 39.3 , 39.5 , 39.7 , 39.9 , 40.1 , 40.3 , 40.6 , 123.2 , 125.7 , 125.8 , 126.2 , 130.2 , 131.6 , 136.2 , 137.0 , 138.3 , 139.0 , 144.0 , 148.8 , 157.0 , 159.4 , 165.0; HRMS calculated 468.3260 (C 29 H 26 D8FN3O H+) Found 468.3264.

[0089] Example 14 (±) 4-Hydroxy-4-[D]-(4'-fluoro(phenyl)-(E)-retinamide (Compound 18) Compound 18 was synthesized using 4-oxo-(4'-fluoro(phenyl)-(E)-retinamide (compound 16) (2 g, 4.91 mmol) and sodium borodeuteride (0.28 g, 5.89 mmol) by following general method B. Yellow solid 1.90 g (94%), mp 173-174 °C. 11H NMR (400 MHz, DMSO-d6) δ ppm 0.99 (s, 3 H) 1.02 (s, 3 H) 1.28 - 1.38 (m, 1 H) 1.49 - 1.68 (m, 2 H) 1.68 - 1.81 (m, 4 H) 1.99 (s, 3 H) 2.35 (s, 3 H) 4.64 (s, 1 H) 6.01 (s, 1 H) 6.14 - 6.28 (m, 2 H) 6.31 (d, 1 H) 6.39 (d, 1 H) 7.01 (dd, 1 H) 7.09 - 7.19 (m, 2 H) 7.61 - 7.71 (m, 2 H) 10.07 (s, 1 H); 2 1H NMR (146 MHz, 0.5% DMSO-d6, in DMSO) δ ppm 3.78 (br s, 1 D). 13 13C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.9 , 28.2 , 28.7 , 29.2 , 34.7 , 35.2 , 39.3 , 39.5 , 39.7 , 39.9 , 40.1 , 40.4 , 40.6 , 115.5 , 115.8, 121.1 , 121.1 , 122.9 , 127.8 , 130.3 , 130.9 , 132.3 , 136.3 , 136.6 , 137.9 , 138.6 , 139.5 , 148.9 , 157.1 , 159.5 , 165.1.

[0090] Example 15 4-Oxo-(4'-fluoro(phenyl-2'3'5'6'-D 4 )-(E)-Retinamide (Compound 19) Compound 19 was synthesized using 4-oxo-ATRA (compound 15) (6 g, 19.1 mmol) and 4-fluoroaniline-2,3,5,6-d4 (2.42 g, 21.0 mmol) by following general method A to give 6.60 g (84%) of an orange solid. mp 156-159 °C. 1 H NMR (400 MHz, DMSO-d6) δ ppm 1.17 (s, 6 H) 1.77 (s, 3 H) 1.78 - 1.84 (m, 2 H) 2.04 (s, 3 H) 2.36 (s, 3 H) 2.43 (t, 2 H) 6.05 (s, 1 H) 6.29 - 6.58 (m, 4 H) 7.03 (dd, 1 H) 10.11 (s, 1 H); 2 H NMR (146 MHz,0.5% DMSO-d6, in DMSO) δ ppm 7.14 (br s, 2 D), 7.66 (br s, 2 D). 13 C NMR (101 MHz, DMSO-d6) δ ppm 12.9 , 13.7 , 13.9 , 27.7 , 34.2 , 35.7 , 37.3 , 123.7 , 126.1 , 129.2 , 130.0 , 133.6 , 136.1 , 137.9 , 138.0 , 140.7 , 148.6 , 157.0 , 159.4 , 160.92 , 164.9 , 198.2.

[0091] Example 16 (±) 4-Hydroxy-4-[H]-(4'-fluoro(phenyl-2'3'5'6'-D 4 )-(E)-Retinamide (Compound 20) By following the general method B, compound 20 was synthesized using 4-oxo-(4'-fluoro(phenyl-2'3'5'6'-D4)-(E)-retinamide (compound 19) (2.5 g, 6.07 mmol) and sodium borohydride (0.28 g, 7.29 mmol) to give 2.35 g (94%) of a yellow solid. mp, sintered at 120 °C, melted at 153-156 °C, 1 H NMR (400 MHz, DMSO-d6) δ ppm 0.99 (s, 3 H) 1.02 (s, 3 H) 1.30 - 1.36 (m, 1 H) 1.52 - 1.62 (m, 2 H) 1.75 (s, 4 H) 1.99 (s, 3 H) 2.35 (s, 3 H) 3.81 (d, 1 H) 4.66 (d, 1 H) 6.02 (s, 1 H) 6.14 - 6.27 (m, 2 H) 6.31 (d, 1 H) 6.39 (d, 1 H) 7.01 (dd, 1 H) 10.07 (s, 1 H); 2 H NMR (146 MHz,0.5% DMSO-d6, in DMSO) δ ppm 7.12 (br s, 2 D), 7.65 (br s, 2 D); 13 C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.9 , 28.3 , 28.9 , 29.2 , 34.7 , 35.2 , 39.3 , 39.5 , 39.7 , 39.98 , 40.1 , 40.4 , 40.60 , 68.7 , 123.0 , 127.8 , 130.3 , 130.9 , 132.3 , 136.2 , 136.6, 137.9 , 138.6 , 139.5 , 148.9, 157.0 , 159.4, 165.0.

[0092] Example 17 (±) 4-Hydroxy-4-[D]-(4'-fluoro(phenyl-2'3'5'6'-D 4 )-(E)-Retinamide (Compound 21) By following the general method B, compound 21 was synthesized using 4-oxo-(4'-fluoro(phenyl-2'3'5'6'-D4)-(E)-retinamide (compound 19) (2.5 g, 6.07 mmol) and sodium borodeuteride (0.3 g, 7.9 mmol) to give 2.23 g (87%) of a yellow solid. mp, sintered at 95 °C, melted at 101-103 °C, 1 H NMR (400 MHz, DMSO-d6) δ ppm 0.99 (s, 3 H) 1.02 (s, 3 H) 1.30 - 1.36 (m, 1 H) 1.50 - 1.62 (m, 2 H) 1.70 - 1.77 (m, 4 H) 1.99 (s, 3 H) 2.35 (s, 3 H) 4.64 (br. s., 1 H) 6.02 (s, 1 H) 6.10 - 6.24 (m, 2 H) 6.25 - 6.34 (m, 1 H) 6.39 (d, 1 H) 7.00 (dd, 1 H) 10.08 (s, 1 H); 2 H NMR (146 MHz,3% DMSO-d6, in DMSO) δ ppm 3.73 (br s, 1D) 7.16 (br s, 2 D) 7.68 (br s, 2 D); 13 C NMR (101 MHz, DMSO-d6) δ ppm 13.0 , 13.7 , 18.9 , 28.2 , 28.7 , 28.9 , 29.2 , 34.7 , 35.2 , 123.0 , 127.8 , 130.3, 130.9 , 132.3 , 132.3 , 136.2 , 136.6 , 137.9 , 138.6 , 139.5 , 148.9 , 157.0 , 159.4 , 165.0

[0093] Example 18: Cellular antiproliferative activity against two TNBC cell lines Compounds 3–9 were evaluated over a range of concentrations to investigate their antiproliferative activity against two TNBC cell lines, MDA-MB-231 and MDA-MB-468, using the well-established 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay. 48、55、56 The parent non-deuterated VNLG-152R (compound 2) was used as a positive control. As shown in Figure 8A, the GI of deuterated compounds 3-9 on MDA-MB231 cells was significantly higher than that of deuterated compounds 3-9. 50 The GI values ​​(range 620–830 nM) were significantly higher than those of the parent protio VNLG-152R (compound 2). 50 In contrast, as shown in FIG. 8B, MDA-MB-468 cells were more sensitive to the deuterated compound (GI 50 values: range 190-530 nM) (1.61-2.79 fold). The reason for this difference is not understood at present. Applicant hypothesizes that enhanced diffusion of these deuterated molecules across the cell membrane of this cell line may result in enhanced intracellular concentrations and inhibition of its proliferation. In Figures 8A and 8B, dose-response curves were generated from MTT assays after six days of exposure to different concentrations of compounds. Each point is the average of replicates from three independent experiments. GI 50 Values ​​were determined from dose-response curves by nonlinear regression analysis using GraphPad Prism.

[0094] Additionally, a similar evaluation was performed in which compounds 3-9 were evaluated in the same manner as described above with respect to Figures 8A and 8B. However, compounds 3-9 were further compared to paclitaxel (PTX). As shown in Figures 9A and 9B, the deuterated analogs (compounds 3-9) were better or equipotent compared to non-deuterated VNLG-152R (compound 2) in in vitro anti-proliferative activity against MDA-MB-231 and MDA-MB-468 TNBC cell lines.

[0095] Example 19: Cellular antiproliferative activity against two non-TNBC cell lines Compounds 2, 4, 8, and 9, as well as paclitaxel (PTX), were evaluated over a range of concentrations to investigate their antiproliferative activity against two non-TNBC cell lines, MCF7 and SKBR3, using the well-established 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay. 48、55、56 The parent non-deuterated VNLG-152R (compound 2) was used as a positive control.

[0096] As shown in Table 1 below, the GI of deuterated compounds 4, 8 and 9 on MCF7 cells 50 Values ​​are the GI of the parent Prothio VNLG-152R (compound 2). 50 The values ​​are identical to those of the deuterated compounds. In contrast, SKBR3 cells appear to be sensitive to deuterated compounds compared to the parent non-deuterated VNLG-152R (compound 2). The reason for this difference is not understood at present. The applicant hypothesizes that enhanced diffusion of these deuterated molecules across the cell membrane of this cell line may result in enhanced intracellular concentrations and inhibition of their proliferation. Each data point is the average of replicates from three independent experiments.

[0097] [Table 1]

[0098] Example 20: Effects of VNLG-152R (Compound 2) and its deuterated analogs on Mnk1, eIF4E, peIF4E, cyclin D1, Bcl2 and Bax Applicant previously reported that the antiproliferative effects of Compound 1 and other Mnk1 / 2 degradation inducers in breast cancer cells are due to degradation of Mnk1 / 2 with consequent depletion of peIF4E and modulation of downstream molecular targets. 48、55、56 Therefore, the deuterated analogs were further investigated for their effects on the degradation of Mnk1 and other relevant downstream molecular targets in both MDA-MB-231 and MDA-MB-468 TNBC cells by Western blot analysis.

[0099] Cell culture and Western blot Human breast cancer cell lines, MDA-MB-231, MDA-MB-468, were procured from ATCC (Manassas, VA) and cultured at 37°C and 5% CO2 in the recommended medium supplemented with 10% heat-inactivated standard fetal bovine serum (FBS, GIBCO) and 1% penicillin-streptomycin (10,000U / ml, Life Technologies). Primary antibodies against Mnk1, eIF4E, p-eIF4E, cyclin D1, BCL2, BAX, β-actin, and secondary HRP-conjugated anti-rabbit used in the study were procured from Cell Signaling Technology, USA. Cells were lysed in radioimmunoprecipitation assay (RIPA) buffer supplemented with 1× protease inhibitors (Roche, Indianapolis, IN, USA), phosphatase inhibitors (Thermo Scientific, Waltham, MA, USA), 1 mmol / L EDTA, and 1 mmol / L PMSF (Sigma), and immunoblot analysis was performed as previously described 48、64 .

[0100] Cell proliferation analysis As previously described 51、56Cell proliferation assays were performed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) compound. Briefly, 2500 cells / well (MDA-MB-231 and MDA-MB-468) were seeded in 96-well plates for 24 h and treated with the indicated concentrations of VNLG on days 2 and 5 over a span of 8 days. Growth inhibitory concentrations (GI) were calculated based on nonlinear regression curve fitting using GraphPad prism 5.0 software (La Jolla, CA). 50 ) was calculated.

[0101] result Non-deuterated VNLG-152R (compound 2) and the vehicle DMSO were used as positive and negative controls, respectively. Equal protein concentrations from breast cancer cells treated with different concentrations (5, 10 and 20 μM) of compounds for 24 hours were resolved by SDS-PAGE and Western blot probed with respective protein antibodies. Vehicle-treated cells were included as a control and all blots were reprobed for β-actin for loading control.

[0102] As shown in Figure 11A, in MDA-MB231 cells, VNLG-152R (compound 2) and deuterated analogs significantly and dose-dependently reduced the expression of Mnk1 and peIF4E without any notable effect on the expression of total eIF4E and the housekeeping protein, β-actin. Figure 11A also shows that the compound also caused significant depletion of downstream target, cyclin D1, and induction of apoptosis via significant downregulation of anti-apoptotic Bcl-2 and upregulation of pro-apoptotic Bax. A similar phenomenon was also observed in MDA-MB-468 cells (Figure 11B). As expected, the potency of non-deuterated VNLG-152R (compound 2) was equipotent with deuterated compounds 3-9.

[0103] Example 21: Pharmacokinetic parameters of deuterated compounds 4, 8, and 9 are superior to those of non-deuterated VNLG-152R (compound 2) in mice As described above, the antiproliferative effects and mechanisms of action of deuterated compounds 3-9 were, in most cases, comparable to the biological activity of the parent non-deuterated VNLG-152R (compound 2). Thus, the deuterated compounds met all the criteria of our objective to develop new potent Mnk1 / 2 degradation inducers with improved pharmacokinetics. Thus, although the pharmacokinetics of VNLG-152R (compound 2) have been previously reported, 65 In this study, the plasma pharmacokinetics (PK) of compounds 3-9 were directly compared, i.e., evaluated under the same study conditions as VNLG-152R (compound 2) after oral administration in female CD-1 mice.

[0104] In vivo pharmacokinetic (PK) studies The PK study was conducted by Aragen Life Sciences Prive Limited (Aragen), Hyderabad, India, a Certified Research Organization (CRO). All procedures in the study were in accordance with the guidelines provided by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) published in The Gazette of India dated January 7, 2010. The institute's CPCSEA update number is 1125 / PO / Rc / S / CPCSEA dated February 13, 2014. The study was initiated with the approval of the Institutional Animal Ethics Committee (IAEC) (IAEC approval number: B-011A). Details are given in the supplementary material.

[0105] To minimize the number of mice used for PK studies using a validated LC / MS / MS assay, both individual (discrete) dosing (set no. 1: compound 5, EM = 463.2937 and set no. 2: compound 7, EM = 463.2937) and cassette dosing (set no. 3: compounds 2, 3, and 7, EM = 459.2686, 462.2874, and 466.3125, respectively, and set no. 4: compounds 4, 8, and 9, EM = 460.2749, 464.2999, and 467.3188, respectively) were used. 59、66The four sets tested were designated by the exact molecular weights of the compounds shown above and in FIG.

[0106] All experiments were performed at least in triplicate and are expressed as mean ± SE, where appropriate. Treatments were compared to controls using Student's t-tests using either GraphPad Prism or Sigma Plot. Differences between groups were considered statistically significant at P < 0.05.

[0107] result Plasma PK profiles were obtained after a single oral (PO) administration of the compounds in mice. As shown in Figures 12A-C, the plasma concentration versus time profiles indicate that non-deuterated VNLG-152R (compound 2) as well as the two deuterated compounds 4 and 8 are rapidly cleared from the systemic circulation, as they were undetectable 8 hours after their administration. Also, VNLG-152R (compound 2) had the best C max It is also noteworthy that compounds 4 and 8 showed very low systemic exposure, whereas compounds 5 and 6 showed low systemic exposure and AUC.

[0108] Important PK parameters are shown in Table 2. The total scores (8–26 and overall ranking) in Table 2 are max , T 1 / 2 The C is the sum of the rankings of four PK parameters, including AUC and MRT. In Table 2, the data are presented as the average of values ​​obtained from three female CD-1 mice. max is the maximum observed plasma concentration. max is the time to maximum concentration. T 1 / 2 is the elimination half-life. AUC stands for AUC(0-∞), which is the area under the concentration-time curve extrapolated from the time of administration to infinity. MRT refers to the mean residence time. Numbers in parentheses indicate the rank order from best (1) to worst (8). The total score (8-26 and overall ranking) is C max , T 1 / 2 , which is the sum of the rankings of four PK parameters including AUC and MRT.

[0109] [Table 2]

[0110] The three deuterated compounds 4, 8 and 9, with total scores of 9 (2nd place), 12 (3rd place) and 8 (1st place), respectively, are superior to non-deuterated VNLG-152R (compound 2), with a total score of 17 (4th place). In particular, compared to non-deuterated VNLG-152R (compound 2), compound 16, which only has C4-deuterium, has the longest half-life (T) extended by 8.44 h. 1 / 2 ) (3.61 h vs. 12.05 h), with a prolonged mean residence time (MRT) of 6.04 h (3.02 h vs. 9.06 h). It is unclear why the AUCs of all deuterated compounds were lower than that of non-deuterated VNLG-152R (compound 2).

[0111] It should be noted that deuterium substitution does not necessarily result in slower metabolism. 67 For example, the deuterated form of paroxetine (CTP-347) demonstrated increased metabolism compared to the non-deuterated form. 66 It is also important to mention here that deuteration as a strategy to alter pharmacokinetics requires the importance of understanding the systemic clearance mechanism, knowing the identity of the metabolic enzymes involved, the extent to which they contribute to metabolic clearance, and the extent to which metabolism contributes to systemic clearance. 66、68 .

[0112] Example 22: Antitumor activity of deuterated compounds 4, 8, and 9 is superior to that of non-deuterated VNLG-152R (compound 2) in mice After confirming the improved pharmacokinetic parameters of deuterated compounds 4, 8, and 9, the antitumor activity of these compounds was tested in MDA-MB-231 and MDA-MB468 xenograft models. In both models, each group of mice was treated with either vehicle (control) or compound at 20 mg / kg, PO, 5 days / week, respectively.

[0113] As shown in Figures 13A and 13B, in the MDA-MB-213 model, non-deuterated compound VNLG-152R reduced tumor growth by 87%, deuterated compounds 4 and 8 reduced tumor growth by 95%, and deuterated compound 9 caused 67% tumor regression. *=p<0.0001.

[0114] Similarly, as shown in Figures 13C and 13D, in the MDA-MB-468 model, non-deuterated compound VNLG-152R reduced tumor growth by 81%, deuterated compound 8 reduced tumor growth by 93%, and deuterated compounds 4 and 9 caused tumor regression of 6.6% and 37%, respectively. *=p<0.0001.

[0115] In both the MDA-MB-231 and MDA-MB-468 models, the body weight of mice in each group increased from approximately 19 g to 21.5 g, indicating that the compound was safe (well tolerated) at effective doses, i.e., without obvious host toxicity.

[0116] Example 23: Molecular mechanism study of VNLG-152R To help identify potential new molecular targets for treatment, we examined the molecular mechanism of VNLG-152R after confirming its antitumor effect. 70、71 Label-free proteomic analysis was performed using MDA-MB-231 cells were treated with VNLG-152R at the concentrations shown in Figure 14A and Figure 14B for 24 hours, followed by Western blotting of whole cell lysates.

[0117] These tested revealed that VNLG-152R significantly upregulated (FC>3, FDR adjusted p<0.0005) the E3 ubiquitin-protein ligase, Synoviolin 1 (SYVN1, also known as DER3, HRD1). See Figure 14A and Figure 14B. Furthermore, as shown in Figure 14A and Figure 14B, the enhanced expression of SYVN1 was dose-dependent. Furthermore, dose-dependent Mnk1 depletion, as well as the suppression of its downstream targets peIF4E and cyclin D1, were also observed. See Figure 14B. Further testing of Mnk1 and Mnk2 with VNLG-152R and its deuterated analogs, such as compounds 4, 8 and 9, are potential future test targets.

[0118] Although the present invention has been described with reference to specific embodiments thereof, those skilled in the art will understand that various modifications may be made thereto without departing from the spirit and scope of the invention. The scope of the appended claims is not limited to the specific embodiments described.

[0119] TIFF2025508983000024.tif196162TIFF2025508983000025.tif244164TIFF2025508983000026.tif244165TIFF2025508983000027.tif25116 3TIFF2025508983000028.tif244160TIFF2025508983000029.tif251162TIFF2025508983000030.tif255162TIFF2025508983000031.tif12162

Claims

1. Compound of formula (A): 【Chemistry 1】 (In the formula, R １ ~R ８ is independently H or D, and R １ ~R ８ At least one of is D. or a pharma- ceutically acceptable salt thereof.

2. R ４ , and R ６ ~R ８ The compound according to claim 1, wherein at least one of is D. 【Request 3】 【Chemistry 2】 【Chemistry 3】 【Chemistry 4】 or a pharma- ceutically acceptable salt thereof. 【Request 4】 【Chemistry 5】 【Chemistry 6】 or a pharma- ceutically acceptable salt thereof.

5. 10. A pharmaceutical composition comprising the compound of claim 1 or a pharma- ceutically acceptable salt thereof and a pharma- ceutically acceptable carrier.

6. The compound is 【Chemistry 7】 【Chemistry 8】 【Chemistry 9】 6. The pharmaceutical composition according to claim 5, which is at least one selected from the group consisting of:

7. The compound is 【Chemistry 10】 6. The pharmaceutical composition according to claim 5, which is at least one selected from the group consisting of:

8. 6. The pharmaceutical composition of claim 5, formulated for oral administration.

9. A method for treating a disease, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of claim 1 or a pharma- ceutically acceptable salt thereof; The method, wherein the disease is one of breast cancer, prostate cancer, bladder cancer, pancreatic cancer, hepatocellular carcinoma, benign prostatic hyperplasia, Kennedy's disease, and blood cancer.

10. The compound is 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 or a pharma- ceutically acceptable salt thereof.

11. The compound is 【Chemistry 14】 【Chemistry 15】 or a pharma- ceutically acceptable salt thereof.

12. The method of claim 9, wherein the disease is breast cancer.

13. The method of claim 12, wherein the breast cancer is triple-negative breast cancer.

14. 10. The method of claim 9, wherein the compound is administered orally.

15. Compound of formula (A): 【Chemistry 16】 (In the formula, R １ ~R ８ is independently H or D, and R １ ~R ８ At least one of is D. or a salt thereof, comprising a compound of formula (B): 【Chemistry 17】 or a salt thereof to said compound of formula (A) or said salt thereof.

16. The conversion of the compound of formula (B) The compound of formula (B) is reacted with a compound of formula (C) or a salt thereof, or a compound of formula (D) or a salt thereof: 【Chemistry 18】 16. The method of claim 15, comprising converting

17. The compound of formula (C) or the salt thereof is reacted with a compound of formula (E) or a salt thereof or a compound of formula (F) or a salt thereof: 【Chemistry 19】 20. The method of claim 16, further comprising converting

18. The compound of formula (D) or a salt thereof is reacted with a compound of formula (G) or a salt thereof or a compound of formula (H) or a salt thereof: 【Chemistry 20】 20. The method of claim 16, further comprising converting

19. Compounds of formula (I): 【Chemistry 21】 or a salt thereof, comprising reacting a compound of formula (B): 【Chemical 22】 to said compound of formula (I).

20. The conversion of the compound of formula (B) The compound of formula (B) is converted into a compound of formula (C): 【Chemistry 23】 Converting to The compound of formula (C) is reacted with a compound of formula (E): 【Chemistry 24】 Then convert it to Converting said compound of formula (E) into said compound of formula (I).

20. The method of claim 19, comprising:

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