Starting from 3-methoxyestra-1,3,5(10),16-tetraene-17-carbaldehydes of natural (13β) and epimeric (13α) series, a series of isomeric 3-hydroxy-17-hydroxymethylestra-1,3,5(10)-trienes, including those containing 16α,17α-annulated cyclopropane and cyclohexane ring D', were prepared using the Corey-Chaykovsky and Diels-Alder reactions followed by reduction-demethylation with diisobutylaluminum hydride and hydrogenation. Target compounds showed antiproliferative effects on MCF-7 breast cancer cells to varying degrees superior to that on MCF-10A cells, in low micromolar concentrations. The ERα-mediated luciferase reporter gene assay demonstrated that obtained steroids without an additional carbocycle or with a cyclopropane 16α,17α-annulated carbocycle are effective ERα activators. In this test, steroids of the natural configuration showed high activity at both 10 nM and 100 nM concentrations, whereas 13α-steroids showed a strong dose-dependent effect, surpassing their natural counterparts at a concentration of 100 nM. The 13β-steroid bearing additional 16α,17α-cyclohexane ring had low activity in the test. A simple docking approach using AutoDock Vina was used as a test for a preliminary assessment of the estrogenicity of the compounds. The scope of its applicability and limitations were shown using examples of synthesized molecules.

Data on the influence of solvents, ratios and nature of reagents on the distribution of typical impurities and yields of target products of the reaction of 19-norpregna-1,3,5(10),16-tetraen-20-one derivatives with 4-dimethylamino-, 3,4-dimethoxy-, and 4-bromobenz-aldehydes and 3-pyridinecarbaldehyde were studied and summarized. A mechanism for the formation of 1,3-diketones in the reaction of 17-acetyl-substituted steroids with 4-dimethyl-aminobenzaldehyde in the presence of lithium diisopropylamide in THF was proposed. The Claisen-Schmidt condensation results were shown to be critically affected by the base and the steroid solubility. Optimization possibilities were considered and an improved procedure for the synthesis of new 21-benzylidene-substituted steroids was proposed.

The synthesis of ionic liquids based on 1-butyl-3-methylimidazolium with different anions—Cl–, Br– and CH3C(O)O– was performed. [BMIm]Cl and [BMIm]Br were obtained by direct interaction of the corresponding butyl halide with N-methylimidazole in acetonitrile solution under heating. [BMIm]OAc was obtained by the ion exchange reaction of [BMIm]Cl with silver acetate. It was found that to obtain homogeneous mixtures of wood pulp LS-0 with ionic liquids, its preliminary impregnation with solutions of imidazolium salts in acetonitrile is preferable. The effective formation temperature of the cellulose solution in ionic liquids is 110°C. The most efficient dissolution occurs in [BMIm]Cl, and when [BMIm]OAc is used, the reaction of attachment of imidazolium to the aldehyde group of hexoses occurs.

Progesterone exerts multiple effects in different tissues through nuclear receptors (nPRs) and through membrane receptors (mPRs) of adiponectin and progestin receptor families. The effect of progesterone on the cells through different types of receptors can vary significantly. At the same time, it affects the processes of proliferation and apoptosis in normal and tumor tissues in a dual way, stimulating proliferation and carcinogenesis in some tissues, suppressing them and stimulating cell death in others. In this study, we have shown the presence of high level of mPRβ mRNA and protein in the HepG2 cells of human hepatocellular carcinoma. Expression of other membrane and classical nuclear receptors was not detected. It could imply that mPRβ has an important function in the HepG2 cells. The main goal of the work was to study functions of this protein and mechanisms of its action in human hepatocellular carcinoma cells. Previously, we have identified selective mPRs ligands, compounds LS-01 and LS-02, which do not interact with nuclear receptors. Their employment allows differentiating the effects of progestins mediated by different types of receptors. Effects of progesterone, LS-01, and LS-02 on proliferation and death of HepG2 cells were studied in this work, as well as activating phosphorylation of two kinases, p38 MAPK and JNK, under the action of three steroids. It was shown that all three progestins after 72 h of incubation with the cells suppressed their viability and stimulated appearance of phosphatidylserine on the outer surface of the membranes, which was detected by binding of annexin V, but they did not affect DNA fragmentation of the cell nuclei. Progesterone significantly reduced expression of the proliferation marker genes and stimulated expression of the p21 protein gene, but had a suppressive effect on the expression of some proapoptotic factor genes. All three steroids activated JNK in these cells, but had no effect on the p38 MAPK activity. The effects of progesterone and selective mPRs ligands in HepG2 cells were the same in terms of suppression of proliferation and stimulation of apoptotic changes in outer membranes, therefore, they were mediated through interaction with mPRβ. JNK is a member of the signaling cascade activated in these cells by the studied steroids.

We propose a new approach to the synthesis of 17-pyrazolylandrostanes on the basis of 17β-[3-(dimethylamino)acryloyl]-16α,17α-epoxy-3β-hydroxyandrost-5-ene. This epoxyandrostene was shown to react with hydrazine hydrate, producing 17α-hydrazinyl-3β,16α-dihydroxy-17β-(1 H -pyrazolyl-3-yl)androst-5-ene that was used in reactions with aromatic aldehydes, leading to 17-hydrazones. The treatment of 17β-[3-(dimethylamino)acryloyl]-16α,17α-epoxy-3β-hydroxyandrost-5-ene with phenylhydrazine led to the formation of 3β,16α-dihydroxy-17-(1-phenyl-1 H -pyrazol-5-yl)androst-5-ene. At the same time, the reactions of epoxyandrostene with methylhydrazine or benzylamine proceeded with conservation of the epoxide ring and led to the formation of 16α,17-epoxyandrostanes containing a 1-methyl-1 H -pyrazol-5-yl or 3-(benzylamino)acryloyl substituent at position 17, respectively. The obtained hydrazones exhibited a moderate antiproliferative activity against the MCF-7 breast cancer cell line. The strongest antiproliferative potential was identified in the case of 17α-[(4-bromobenzylidene)hydrazinyl]-3β,16α-dihydroxy-17β-(1 H -pyrazol-3-yl)androst-5-ene with an IC 50 value around 16 μM.

This study aimed to investigate the activities of novel 20(R)-3,20-dihydroxy-19-norpregn-1,3,5(10)-trienes (kuz7 and kuz8b) of natural 13β- and epimeric 13α-series against triple-negative MDA-MB-231 breast cancer cells. High antiproliferative activity of synthesized compounds kuz8b and kuz7 against MDA-MB-231 triple-negative cancer cells was revealed. The steroid kuz7 of natural 13β-configuration was more active against MDA-MB-231 cells than the 13α-steroid kuz8b. Cell cycle analysis revealed common patterns for the action of both tested compounds. The number of cells in the subG1 phase increased in a dose-dependent manner, indicating induction of apoptosis, which was also verified by PARP cleavage. In contrast, the number of cells in the G0/G1 phase decreases with increasing compound concentration. Steroid kuz7 at micromolar concentrations reduced the expression of GLUT1, a glucose transporter. High efficacy of the combination of kuz7 with biguanide metformin was shown, and synergistic effects on MDA-MB-231 cell growth and expression of the anti-apoptotic protein Bcl-2 were revealed. According to the obtained results, including the high activity of kuz7 against triple-negative cancer cells, the detected induction of apoptosis, and the decrease in GLUT1 expression, 13β-steroid kuz7 is of interest for further preclinical studies both alone and in combination with the metabolic drug metformin.

In this paper, we consider the studies carried out by the authors aimed at developing new hybrid structures and methods for obtaining a family of photochromic labels capable of photocontrolled interaction with inorganic components, as well as the results of studying their photochromic behavior and the selectivity of complex formation processes. In this paper, special attention is devoted to the choice of the desired composition and structure of label molecules and the practical implementation of laboratory technology for the synthesis of a set of target compounds with specified optical parameters. The data obtained open up prospects for the use of a new generation of photochromes based on functionalized spiropyrans as new hybrid materials for the creation on their basis: metal detectors, components of photochromic systems, and prototypes of molecular electronics smart devices.

Methods for extending the side chain of 20-oxopregnanes were summarized. The possibilities of alkylation of 3-methoxy-19-norpregna-1,3,5(10)-trien-20-one and its 16α,17α-methano and -butano derivatives with alkyl halides in the presence of lithium diisopropylamide were studied. The Claisen—Schmidt condensation of these steroids with aromatic aldehydes resulted in a series of the corresponding 21-benzylidenesteroids. Carrying out the Claisen—Schmidt condensation in an aprotic solvent in the presence of lithium diisopropylamide yielded, in addition to the target 21-benzylidenesteroids, the corresponding 21-benzoyl derivatives — steroidal 1,3-diketones.

A new approach to the synthesis of isomeric 19-hydroxypregn-4-en-20-one (1) and 19-hydroxy-5β-pregn-3-en-20-one (2), the selective ligands of membrane progesterone receptors, in a ratio of 1.5:1 is presented. The effect of these steroids on the gene expression of several cytokines (TNFα, IL-1β, IL-6, and TGFβ) in K562 cells was studied. Progesterone and compound 1 acted unidirectionally, significantly decreasing the TGFβ mRNA levels in these cells. The highest activity was exhibited by compound 2 which regulated the expression of the three studied genes, increasing the level of TNFα mRNA and IL-1β mRNA and decreasing the level of TGFβ mRNA. The immunomodulatory effect of the synthesized selective ligands of membrane progesterone receptors found in K562 cells is comparable (for 1) or even surpasses (for 2) the effect of a natural hormone.

The monograph summarizes the information over the past 20 years on the currently widely used and promising methods for the synthesis of estra-1,3,5(10)-triene derivatives by modifying natural estrogens - estrone and estradiol. The main practical goals of modifying this class of steroids and achievements in the chemistry of steroidal antiestrogens, which are promising drugs for hormonal therapy, are considered. Special attention is paid to the stereochemical features of the reactions and the specific problems of modification of the steroid nucleus of estratrienes associated with the presence of an aromatic fragment in their structure. In addition, the data on the reactivity and stereochemical aspects of the transformations of 13-epiestratriene steroids were summarized. The monograph is intended for a wide range of specialists in the field of organic synthesis, organic, bioorganic, and medicinal chemistry.

Triple-negative cancer is characterized by a lack of expression of estrogen receptor α (ERα), progesterone receptor, and HER2/neu, which determines certain difficulties in the treatment of this molecular subtype of cancer. The development of drug candidates for triple-negative breast cancer is especially relevant. The work aims to design new type pentaranes (3-hydroxy-17(1-hydroxyalkyl(aryl))-16,17-cycloalkano)-estra-1,3,5(10)-trienes) and to evaluate their biological activity against breast cancer cells.

Progesterone and its synthetic analogues act on cells through different types of receptors, affecting proliferation and apoptosis. These compounds exert their effect through the nuclear receptors and the insufficiently studied membrane progesterone receptors (mPRs) belonging to the progestin and adiponectin Q receptor (PAQR) family. We have identified two selective ligands of mPRs that activate only this type of progesterone receptors – 19-hydroxypregn-4-en-20-one (LS-01) and 19-hydroxy-5β-pregn-3-en-20-one (LS-02). The goal of this work is to study the effect of these compounds on proliferation and death of human pancreatic adenocarcinoma cells BxPC3 and involvement of the two kinases (p38 MAPK and JNK) in signaling pathways activated by progestins through mPRs. It was shown that progesterone and the compound LS-01 significantly ( p < 0.05) inhibited the BxPC3 cell viability, with JNK serving as a mediator. The identified targets of these two steroids are the genes of the proteins Ki67, cyclin D1, PCNA, and p21. Progesterone and the compound LS-01 significantly ( p < 0.05) stimulate DNA fragmentation, enhancing the cell death. The p38 mitogen-activated protein kinase (MAPK) is a key mediator of this process. The BCL2A1 protein gene was identified as a target of both steroids. The compound LS-02 significantly ( p < 0.05) alters membrane permeability and changes the exposure of phosphatidylserine on the outer membrane leaflet, also enhancing the cell death. This compound acts on these processes by activating both kinases, JNK and p38 MAPK. The compound LS-02 targets the genes encoding the proteins HRK, caspase 9, and DAPK.

A directed synthesis of two progesterone metabolites, allopregnanolone and 3a-hydroxy-pregn-4-en-20-one, from Δ16-pregnanolone and progesterone, respectively, was carried out by a reduction of the carbonyl groups in positions 3 and subsequent inversion of the configuration of the resulting alcohols by the Mitsunobu reaction. The selectivity of the reduction of the conjugated carbonyl group in position 3 of progesterone with sodium borohydride in the presence of cerium(III) chloride (Luche reduction) was demonstrated. The ef ect of the obtained metabolites on the proliferation of breast cancer cells of the MCF-7 and T47D lines under normal and steroid-free conditions was studied. It is shown that the ef ect of these compounds on the proliferation depends on the presence of additional steroids in the culture medium. Metabolites exerted small cytostatic ef ects on the growth of the MCF-7 cells under standard conditions, while the transfer of the cells to a steroid-free medium weakened these cytotoxic ef ects. In the experiments with the T47D line cells, the cell growth was stimulated under both standard and steroid-free conditions. Allopregnanolone and progesterone stimulate the growth to a greater extent under steroid-free conditions than under standard ones.

Breast cancer is the most common cancer in women. There is an urgent need for new drugs with novel modes of effect on tumors. It is very important to develop compounds with high activity against tumor cells and low toxicity on normal epithelium. The development of new anti-tumor drugs based on steroidal compounds is a promising approach for obtaining agents that combine antiestrogenic, cytostatic and proapoptotic effects. The aim of the work is to study the mechanism of anticancer action of new steroidal compound, 3,20 (R)-dihydroxy-19-norpregnatriene (Kuz7). MCF-7 breast cancer cell line and MCF-10A human mammary epithelial cell line were obtained from the ATCC collection. Antiproliferative activity was measured by MTT. ERα activity was assessed by gene-reporter assay. Protein expression was measured by standard immunoblotting. We have developed a new type of anticancer steroids - a series of 3-hydroxyestra-1,3,5(10)-trienes of natural and epimeric 13α-configuration with 17th-side chain bearing the second hydroxy group and 16,17-fused three- or six-membered carbocycle (or without it). Most of these compounds exhibited cytotoxic and ERα inhibiting activities on MCF-7 cells and have low toxicity in the MCF-10A normal epithelial cells. 3,20(R)-dihydroxy-19-norpregnatriene (Kuz7) was selected as a leader molecule for in-depth study. At low doses, Kuz7 caused significant accumulation of p21 and inhibition of CDK2 and CDK4 in MCF-7 cells. Apoptosis provoked by Kuz7 was determined by fragmented PARP. Compound kuz7 is considered as an effective inhibitor of proliferation and ERα signaling in MCF-7 cells. kuz7 decreased cyclin D1 expression and caused cell arrest in G0 phase. New series of highly active antitumor steroids has been developed. The lead compound, kuz7, blocks the growth of MCF-7 cells through induction of apoptosis and regulation of hormonal pathways and cell cycle. Further studies, in vitro and in vivo, are needed to confirm our findings in relation to luminal type A breast cancer. The work was supported by RFBR (project 19-03-00246).

A convenient preparative synthesis of 3-methoxyestra-1,3,5(10),16-tetraene-17-carb-aldehyde and its epimeric 13α-analog, the key structures in the synthesis of steroid estrogen receptor modulators and potential antitumor agents, was developed starting from estrone and including the generation of intermediate 16-dehydro-17-carbonitriles and their reaction with diisobutylaluminum hydride. The influence of conformational dif erences in the steroid molecules with the natural 13β- and epimeric 13α-configurations on the reactivity of intermediate compounds and the stereoselectivity of the trimethylsilyl cyanide addition to the carbonyl group of methyl ethers of epimeric estrones was considered.