Phytoecdysones and their derivatives for use in the treatment of respiratory dysfunction

Abstract

The present invention relates to phytoecdysones and derivatives thereof for use in the treatment of respiratory dysfunction in mammals, in particular as part of neuromuscular diseases, and more particularly when said respiratory dysfunction is associated with a decrease in the mechanical properties of lung tissue.

Description

Technical Field

[0001] The present invention relates to the use of phytoecdysones, and semi-synthetic derivatives of phytoecdysones, for the treatment of respiratory dysfunction, particularly respiratory dysfunction as part of neuromuscular diseases.

Background Art

[0002] Neuromuscular diseases are characterized by functional changes in the motor unit composed of motor neurons, neuromuscular junctions, and skeletal muscles. In addition to the motor dysfunction of patients due to these pathologies, a very large number of acute or progressive neuromuscular diseases cause respiratory muscle insufficiency, which can lead to respiratory failure, pneumonia, and death of the patient. In fact, respiratory disorders are the main cause of death in patients with neurological diseases (Non-Patent Document 1).

[0003] Patients with neuromuscular diseases may develop respiratory dysfunction, which may manifest clearly with frequent occurrences of infectious diseases such as pneumonia or bronchitis (mainly due to the lack of coughing effect), a feeling of shortness of breath, and difficulty expectorating. However, in some cases, the symptoms are not so obvious, and the patient has loss of appetite, headache, sweating, or severe fatigue accompanied by significant weight loss. Therefore, it is important to detect respiratory diseases as early as possible.

[0004] Care for respiratory problems in patients with neuromuscular diseases has improved significantly in recent years, making it possible to extend the lifespan of patients such as children with Duchenne muscular dystrophy. Respiratory parameters are regularly monitored using clinical tests, spirometry imaging to assess respiratory function, or gas measurements (measurement of oxygen and carbon dioxide levels in arterial blood) to evaluate the quality of gas exchange. These regular tests can detect the consequences of muscle weakness and pulmonary dysfunction, thereby allowing for the adaptation of medical care for these patients to compensate for their reduced respiratory function and improve their quality of life (Non-patent documents 2-4). The above care can be provided at several levels: it may be aimed at maintaining the mobility and flexibility of the respiratory system (respiratory physiotherapy by active or passive mobilization or by mechanical hypersuction); or it may be aimed at clearing the airways to remove secretions produced by the bronchi (coughing assistance or bronchial drainage); or finally, when spontaneous breathing no longer meets the body's needs, it may be aimed at supplementing the patient's breathing by non-invasive ventilation, or in the most severe cases, by ventilation via tracheostomy.

[0005] Damage to the cortex, brainstem, spinal cord, motor neurons, peripheral nerves, neuromuscular junctions, or muscles can all lead to respiratory problems.

[0006] There are numerous causes of chronic muscle disorders that lead to respiratory muscle dysfunction, including (congenital, hereditary, or acquired) myopathy, myasthenia gravis, or myotonia.

[0007] For example, Duchenne muscular dystrophy (DMD), a respiratory failure leading to numerous pulmonary complications, is the cause of the majority of deaths observed in DMD patients (Non-Patent Literature 5-9). DMD is the most common form of muscular dystrophy. It affects 1 in 3,500 boys and is caused by a mutation affecting the dystrophin gene on the X chromosome. Becker muscular dystrophy (BMD), a relatively less severe form, also involves the dystrophin gene and affects 1 in 18,000 boys. Boys with DMD generally do not experience difficulty breathing or coughing as long as they are still able to walk. As they age and the disease affects the respiratory muscles, affected boys are at risk of respiratory infections, often due to the ineffectiveness of coughing. Smooth muscle cells, and by extension the smooth muscles of the airways, are involved in numerous respiratory diseases.

[0008] In addition to the myopathic process, abnormalities in the pulmonary system (airways or lungs) itself are strongly involved in respiratory failure in DMD patients (Non-Patent Literature 10). In fact, lung compliance (the lung's ability to change its volume in response to pressure changes) is reduced mainly for two reasons: alveolar contraction and collapse caused by hypopnea (atelectasis); and the development and obstruction of airway fibrosis, which results in increased airway resistance (Non-Patent Literature 11).

[0009] In patients with muscular dystrophy, the elastic properties of the lungs are impaired, and lung extensibility is reduced. The cause of this reduced lung extensibility in muscular dystrophy is still not fully understood. Nevertheless, various hypotheses have been proposed in an attempt to explain the reduced lung elasticity, including: incomplete maturation of lung tissue in the context of congenital disease; atelectasis induced by hypopnea; increased tension on the surface of the alveoli; or damage to the lung parenchyma due to fibrosis. Furthermore, one of the important factors for explaining such reduced extensibility and lung compliance is low lung volume respiration, a characteristic of muscular dystrophy (Non-Patent Literature 11).

[0010] Another mechanism involved is the chronic and progressive deterioration of the respiratory muscles, which effectively limits the range of lung activity. In fact, part of the elastic properties of a system are determined by the stresses it experiences. Total lung capacity is the result of the balance between the instantaneous elastic contraction pressure during respiration and the pressure generated by the contraction of the inspiratory muscles. When the latter decreases, total lung capacity also decreases as a result, which alters expiratory parameters and leads to decreased lung compliance.

[0011] In conclusion, respiratory distress characterized by the respiratory system's inability to provide adequate oxygen intake and carbon dioxide elimination is common to patients with DMD.

[0012] Therefore, evaluating respiratory system dysfunction in the MDX mouse, the most widely used mouse model of Duchenne muscular dystrophy, is an important parameter to consider when establishing and evaluating therapeutic solutions in the context of neuromuscular diseases.

[0013] Evaluating the respiratory system of MDX mice during preclinical trials offers several significant advantages: on the one hand, this evaluation is relevant to clinically significant deficiencies; on the other hand, spirometry or plethysmography measurements are non-invasive, can be repeated longitudinally during the trial, and can optionally be used as a criterion for evaluating the effectiveness of various treatments for muscular dystrophy.

[0014] Various groups have shown interest in the respiratory function of MDX mice compared to healthy control mice and have reported respiratory dysfunction in MDX mice (Non-Patent Literature 12-15). While there have been some variations in the severity of the dysfunction and the age of onset, all of these reports describe changes in respiratory parameters under normal oxygen conditions (Non-Patent Literature 16) or in response to hypercapnia (Non-Patent Literature 13). [Prior art documents] [Non-patent literature]

[0015]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Non-Patent Document 7

Non-Patent Document 8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

[2003]

Non-Patent Document 13

[2003]

Non-Patent Document 14

Non-Patent Document 15

[2007]

Non-Patent Document 16

Summary of the Invention

Problems to be Solved by the Invention

[0016] The inventors have discovered that plant ecdysones, and semi-synthetic derivatives of plant ecdysones, significantly improve the respiratory function of mammals with neuromuscular diseases by restricting the time-dependent changes in respiratory parameters and by improving the mechanical parameters of the respiratory system. The respiratory parameters, and the mechanical parameters of the respiratory system, are determined, respectively, by whole body plethysmography of awake animals and by a piston ventilator controlled by a central control unit (commonly referred to as a "computer") in anesthetized animals, the ventilator using forced oscillation techniques such as a device known as FlexiVent™. These effects indicate an improvement in the respiratory function of mammals with hereditary or acquired neuromuscular diseases.

[0017] Plant ecdysones are an important family of plant polyhydroxylated steroids that are structurally related to the insect molting hormone. These molecules are produced by a number of plant species and are involved in their defense against pests. Most plant ecdysones are 20-hydroxyecdysone.

Means for Solving the Problems

[0018] For this purpose, the present invention relates to at least one plant ecdysone, and / or at least one semi-synthetic derivative of a plant ecdysone, for use in the treatment of respiratory dysfunction.

[0019] The present invention preferably relates to a composition comprising at least one plant ecdysone and / or at least one semi-synthetic derivative of a plant ecdysone for use in the treatment of respiratory dysfunction. <0**********> In certain embodiments, the present invention also meets the following characteristics, implemented separately or in any technically possible combination.

[0021] The plant ecdysones and their derivatives are advantageously purified to pharmaceutical grade.

[0022] A plant ecdysone usable according to the present invention is, for example, 20-hydroxyecdysone, and a semi-synthetic derivative of plant ecdysone usable is, for example, a semi-synthetic derivative of 20-hydroxyecdysone.

[0023] For this purpose, according to one embodiment, the composition comprises 20-hydroxyecdysone and / or at least one semi-synthetic derivative of 20-hydroxyecdysone.

[0024] 20-hydroxyecdysone and its derivatives are advantageously purified to pharmaceutical grade.

[0025] The 20-hydroxyecdysone used is more preferably in the form of a plant extract rich in 20-hydroxyecdysone, or a composition containing 20-hydroxyecdysone as an active ingredient. Examples of plant extracts rich in 20-hydroxyecdysone include extracts of Stemmacantha carthamoides (also known as Leuzea carthamoides), Cyanotis arachnoidea, and Cyanotis vaga.

[0026] The resulting extract is preferably purified to pharmaceutical grade.

[0027] In one embodiment, 20-hydroxyecdysone is in the form of a plant extract or a part of a plant, the plant being selected from plants containing at least 0.5% of 20-hydroxyecdysone by the dry weight of the plant, and the extract containing at least 95%, preferably at least 97%, of 20-hydroxyecdysone. The extract is preferably purified to pharmaceutical grade.

[0028] The above extract will henceforth be referred to as BIO101. It is characterized by containing 0-0.05% of the dry weight of the above extract of trace compounds and other impurities that may affect the safety, usability, or efficacy of the above extract in pharmaceutical applications.

[0029] According to one embodiment of the present invention, the impurity is a compound containing 19 or 21 carbon atoms, such as rubrosterone, dihydrolubrosterone, or poststerone.

[0030] The plants used as the source for BIO101 production are preferably selected from Stemmacantha carthamoides (also known as Leuzea carthamoides), Cyanotis arachnoidea, and Cyanotis vaga.

[0031] Plant ecdysone derivatives, particularly 20-hydroxyecdysone derivatives, can be obtained by semi-synthesis, and in particular in the form described in European Patent Application No. 15732785.9.

[0032] According to one preferred embodiment, the present invention relates to a composition for use in mammals in the treatment of respiratory dysfunction, more particularly in acquired hereditary neuromuscular diseases, such as neuromuscular diseases of motor neurons, and / or neuromuscular junctions, and / or striated muscles.

[0033] According to a particular embodiment, the present invention relates to a composition for use in mammals in the treatment of respiratory dysfunction associated with striated muscle and / or smooth muscle disorders.

[0034] In certain embodiments, the present invention relates to a composition for use in mammals in the treatment of respiratory dysfunction caused at least partially by smooth muscle damage.

[0035] According to a particular embodiment, the present invention relates to a composition for use in mammals in the treatment of respiratory dysfunction associated with bronchial hyperreaction.

[0036] In one embodiment, the present invention relates to the composition for use in mammals in the treatment of respiratory disorders in which the bronchial hyperreaction is related to the function of bronchial smooth muscle.

[0037] In one embodiment, the present invention relates to a composition for use in mammals in the treatment of respiratory dysfunction associated with at least one of the respiratory parameters selected from Penh value, peak inspiratory flow, peak expiratory flow, relaxation time, and respiratory rate. The composition advantageously reduces the conditions of these respiratory parameters.

[0038] In one embodiment, the present invention relates to the composition for use in mammals in the treatment of respiratory dysfunction related to at least one of the mechanical parameters of lung tissue. The mechanical parameters of the lung tissue are lung elastance, compliance, and resistance.

[0039] In certain embodiments, the present invention relates to the above-mentioned composition for use in mammals in the treatment of respiratory disorders associated with decreased lung compliance and / or increased lung resistance and / or decreased lung elastance.

[0040] In certain embodiments, the present invention relates to compositions for use in mammals in the treatment of diseases in which respiratory dysfunction is associated with alveolar contraction and collapse and / or the development of fibrosis.

[0041] In certain embodiments, plant ecdysone is administered to humans at doses of 3–15 mg / kg / day. “Plant ecdysone” is understood here to mean common plant ecdysone and its derivatives, and 20-hydroxyecdysone and its derivatives (particularly in extract form).

[0042] Preferably, plant ecdysone is administered to adult humans in a dose of 200-1,000 mg / day in one or more divided doses, and to human children or infants in a dose of 5-350 mg / day in one or more divided doses. "Plant ecdysone" is understood here to mean general plant ecdysone and its derivatives, and 20-hydroxyecdysone and its derivatives (especially in extract form).

[0043] In some embodiments, the composition comprises at least one compound that is considered to be a derivative of a plant ecdysone, wherein the at least one compound is of general formula (I):

[0044] [ka]

[0045] This is a compound where V-U is a carbon-carbon single bond, Y is a hydroxyl group or hydrogen, or V-U is an ethylenically charged C=C bond; X is oxygen; Q is a carbonyl group; R 1 The group is selected from: (C1-C6)W(C1-C6) group; (C1-C6)W(C1-C6)W(C1-C6) group; (C1-C6)W(C1-C6)CO2(C1-C6) group; (C1-C6)A group (where A represents a heterocycle arbitrarily substituted with a group of the type OH, MeO, (C1-C6), N(C1-C6), CO2(C1-C6)); CH2Br group; W is a heteroatom selected from N, O, and S, preferably O, and more preferably S.

[0046] In the context of the present invention, "(C1-C6)" means any linear or branched alkyl group having 1 to 6 carbon atoms, particularly methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, or n-hexyl groups. Advantageously, this involves methyl, ethyl, isopropyl, or t-butyl groups, particularly methyl or ethyl groups, and more specifically, methyl groups.

[0047] In one preferred embodiment, in formula (I): Y is a hydroxyl group; R 1a is selected from: (C1-C6)W(C1-C6) group; (C1-C6)W(C1-C6)W(C1-C6) group; (C1-C6)W(C1-C6)CO2(C1-C6) group; (C1-C6)A group, (where A represents a heterocycle arbitrarily substituted with groups of the type OH, MeO, (C1-C6), N(C1-C6), CO2(C1-C6)); W is a heteroatom selected from N, O, and S, preferably O, and more preferably S.

[0048] In some embodiments, the above composition comprises at least one compound selected from the following compounds: No. 1: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-10,13-dimethyl-17-(2-morpholinoacetyl)-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthrene-6-one; No.2: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-17-[2-(3-hydroxypyrrolidine-1-yl)acetyl]-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthrene-6-one; No.3: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-17-[2-(4-hydroxy-1-piperidyl)acetyl]-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthrene-6-one; No.4: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-17-[2-[4-(2-hydroxyethyl)-1-piperidyl]acetyl]-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthrene-6-one; No. 5: (2S,3R,5R,10R,13R,14S,17S)-17-[2-(3-dimethylaminopropyl(methyl)amino)acetyl]-2,3,14-trihydroxy-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthrene-6-one; No.6: 2-[2-oxo-2-[(2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-10,13-dimethyl-6-oxo-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-17-yl]ethyl]ethylsulfanyl acetate; No. 7: (2S,3R,5R,10R,13R,14S,17S)-17-(2-ethylsulfanylacetyl)-2,3,14-trihydroxy-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthrene-6-one; No. 8: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-17-[2-(2-hydroxyethylsulfanyl)acetyl]-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1Hcyclopenta[a]phenanthren-6-one.

[0049] In some embodiments, the composition comprises at least one compound that is considered to be a derivative of a plant ecdysone, wherein the at least one compound is of general formula (II):

[0050] [ka]

[0051] The compound represented by formula (II) will henceforth be referred to as BIO103.

[0052] In some embodiments, the above composition is incorporated into an orally administered, pharmaceutically acceptable formulation.

[0053] In the context of the present invention, "pharmaceutically acceptable" generally means that the pharmaceutical composition is safe, non-toxic, and useful for preparing pharmaceutical compositions that are acceptable for veterinary and human pharmaceutical use.

[0054] The present invention will be better understood by referring to the drawings and reading the following description, which is given as a non-limiting example. [Brief explanation of the drawing]

[0055] [Figure 1A] Figure 1A shows the curves of Penh values ​​in response to increasing methacholine dose, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles) and mdx mice (n=23, white squares) before treatment (D0: day 0), where ** represents p<0.001 and *** represents p<0.0001. Hereafter, n corresponds to the sample size, and p corresponds to the "p-value" used to quantify the statistical significance of the results, where * represents p<0.05, ** represents p<0.001, *** represents p<0.0001, and ns represents not significant difference. [Figure 1B] Figure 1B shows the curves of peak inspiratory flow in healthy control mice C57Black10 (n=12, white circles) and mdx mice (n=23, white squares) before the start of the study, measured by plethysmography, in response to increasing methacholine dose, where * represents p<0.05 and ** represents p<0.001. [Figure 1C] Figure 1C shows the curves of peak expiratory flow in response to increasing methacholine dose, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles) and mdx mice (n=23, white squares) before treatment (D0: day 0), where ** represents p<0.001. [Figure 1D]Figure 1D shows the curves of relaxation time in response to increasing methacholine dose, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles) and mdx mice (n=23, white squares) before treatment (D0: day 0), where ** represents p<0.001 and *** represents p<0.0001. [Figure 1E] Figure 1B shows the respiratory rate curves in response to increasing methacholine dose, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles) and mdx mice (n=23, white squares) before treatment (D0: day 0), where * indicates p<0.05 and *** indicates p<0.0001. [Figure 2A] Figure 2A shows curves of Penh values ​​in response to increasing methacholine dose, measured by plethysmography, before study initiation (D0), after randomizing animals into three different groups, where ** is p<0.001 and *** is p<0.0001: healthy control mice C57Black10 (n=12, white circles); untreated mdx mice (n=11, white squares, "mdx" group); and mdx mice that would be treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 2B] Figure 2B shows the curves of peak inspiratory flow in response to increasing methacholine dose, measured by plethysmography, before the start of the study (D0), after randomizing animals into three different groups: healthy control mice C57Black10 (n=12, white circles); untreated mdx mice (n=11, white squares, "mdx" group); and mdx mice that would be treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 2C]Figure 2C shows the curves of peak expiratory flow in response to increasing methacholine dose, measured by plethysmography, before the start of the study (D0), after randomizing animals into three different groups: healthy control mice C57Black10 (n=12, white circles); untreated mdx mice (n=11, white squares, "mdx" group); and mdx mice that would be treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 2D] Figure 2D shows the curves of relaxation time in response to increasing methacholine dose, measured by plethysmography, before the start of the study (D0), after randomizing animals into three different groups: healthy control mice C57Black10 (n=12, white circles); untreated mdx mice (n=11, white squares, "mdx" group); and mdx mice that would be treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 2E] Figure 2E shows the curves of respiratory rate in response to increasing methacholine dose, measured by plethysmography, before the start of the study (D0), after randomizing animals into three different groups: healthy control mice C57Black10 (n=12, white circles); untreated mdx mice (n=11, white squares, "mdx" group); and mdx mice that would be treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 3A] Figure 3A shows curves of Penh values ​​in response to increasing methacholine dose 30 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group), where ** represents p<0.001. [Figure 3B]Figure 3B shows the curves of peak inspiratory flow in response to increasing methacholine dose 30 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group), where * represents p<0.05 and ** represents p<0.001. [Figure 3C] Figure 3C shows the curves of peak expiratory flow in response to increasing methacholine dose 30 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group), where ** represents p<0.05. [Figure 3D] Figure 3D shows the curves of relaxation time in response to increasing methacholine dose 30 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group), where * represents p<0.05. [Figure 3E] Figure 3E shows the respiratory rate curves in response to increasing methacholine dose 30 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 4A]Figure 4A shows curves of Penh values ​​60 days after the start of treatment in response to increasing methacholine dose for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group), as measured by plethysmography, where *** represents p<0.0001. [Figure 4B] Figure 4B shows the curves of peak inspiratory flow in response to increasing methacholine dose 60 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group), where * represents p<0.05. [Figure 4C] Figure 4C shows the curves of peak expiratory flow in response to increasing methacholine dose 30 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 4D] Figure 4D shows the curves of relaxation time in response to increasing methacholine dose 60 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 4E]Figure 4E shows the respiratory rate curves in response to increasing methacholine dose 60 days after the start of treatment, measured by plethysmography, for healthy control mice C57Black10 (n=12, white circles), untreated mdx mice (n=11, white squares, "mdx" group), and mdx mice treated with BIO101 (n=12, black squares, "mdx BIO101" group). [Figure 5A] Figure 5A shows curves of changes in Penh values ​​in healthy mice C57Black10 at baseline (n=12, black circles, D0) and end of the study (n=12, white circles, D60) as the dose of metacholine was increased, as measured by plethysmography. [Figure 5B] Figure 5B shows curves of change in Penh values ​​in untreated mdx mice at baseline (n=23, black circles, D0) and end of the study (n=12, white circles, D60) as the dose of metacholine increased, as measured by plethysmography, where *** represents p<0.0001. [Figure 6A] Figure 6A shows Penh values ​​measured by plethysmography for healthy C57Black10 mice (n=12) and untreated mdx mice (n=23) at baseline (D0) with a methacholine dose of 40 mg / ml, where *** represents p<0.0001. [Figure 6B] Figure 6B shows Penh values ​​measured by plethysmography for healthy C57Black10 mice (n=12), untreated mdx mice (n=12), and mdx mice treated with BIO101 (n=12) at 60 days (D60) of the study, with a methacholine dose of 40 mg / ml, where *** represents p<0.0001. [Figure 7A]Figure 7A shows the changes in lung resistance in healthy C57Black10 mice (n=10), untreated mdx mice (n=10), and mdx mice treated with BIO101 (n=10) as the dose of metacholine was increased, as measured by a piston ventilator (FlexiVent®), where ** represents p<0.001 and *** represents p<0.0001. [Figure 7B] Figure 7B shows the changes in lung compliance in healthy C57Black10 mice (n=10), untreated mdx mice (n=10), and BIO101-treated mdx mice (n=10) as the dose of metacholine was increased, as measured by piston ventilators, where *** represents p<0.0001. [Figure 7C] Figure 7C shows the changes in lung elastance in healthy C57Black10 mice (n=10), untreated mdx mice (n=10), and mdx mice treated with BIO101 (n=10) as the dose of metacholine was increased, as measured by piston ventilators, where *** represents p<0.0001. [Figure 8A] Figure 8A shows lung resistance values ​​for healthy C57Black10 mice (n=10), untreated mdx mice (n=10), and mdx mice treated with BIO101 (n=10) 60 days after the start of the study, measured by piston ventilators, at a methacholine dose of 20 mg / ml, where *** represents p<0.0001. [Figure 8B] Figure 8B shows lung compliance values ​​for healthy C57Black10 mice (n=10), untreated mdx mice (n=10), and mdx mice treated with BIO101 (n=10) 60 days after the start of the study, measured by piston ventilators, at a methacholine dose of 20 mg / ml, where *** represents p<0.0001. [Figure 8C]Figure 8C shows the lung elastance values ​​for healthy C57Black10 mice (n=10), untreated mdx mice (n=10), and mdx mice treated with BIO101 (n=10) 60 days after the start of the study, measured by piston ventilators, at a methacholine dose of 20 mg / ml, where ** represents p<0.05 and ** represents p<0.001. [Modes for carrying out the invention]

[0056] The present invention will now be described in specific contexts of some of its preferred, non-limiting fields of application.

[0057] 1. Purification method for BIO101 BIO101 is prepared from 90% pure 20-hydroxyecdysone according to the following steps: i) Dissolve 90% pure 20-hydroxyecdysone in methanol using heat, then filter and partially concentrate it; ii) Add three times the amount of acetone; iii) Cooling while stirring to a temperature of 0-5°C; iv) A step of filtering the obtained precipitate; v) A step of rinsing continuously with acetone and water; and vi) Drying step.

[0058] This purification involves a recrystallization process suitable for this molecule and that can be carried out on an industrial scale.

[0059] Filtration in step i) is performed using a 0.2 μm particle filter.

[0060] The partial enrichment in step i) is advantageously carried out by vacuum distillation at a temperature of about 50°C in the presence of MeOH.

[0061] The drying step (vi) is carried out under vacuum at a temperature of approximately 50°C.

[0062] 2. Bioactivity of BIO101 We used 12-week-old male mice C57BL / 10ScSnJ (healthy mice shown as "C57Black10" in the diagram) and C57BL / 10ScSn-Dmdmdx / J (a mouse model of Duchenne muscular dystrophy shown as "mdx" in the diagram). These mice were divided into three groups of 12 each: an untreated group of healthy control mice C57Black10; an untreated group of mdx mice (mdx); and a group of mice regularly treated orally with a dose of 50 mg / kg / day using drinking water.

[0063] The respiratory function of all mice was assessed by whole-body plethysmography before the start of the study (D0), 30 days after the start of treatment (D30), and 60 days after the start of treatment (D60). After two months of treatment, respiratory function was non-invasively assessed using a piston ventilator controlled by a central control unit (commonly referred to as the "computer") immediately before the animals were killed. The ventilator used forced vibration techniques, such as a device known as FlexiVent®, to more directly determine the mechanical parameters of the respiratory system.

[0064] a. Plastimographic analysis of the effects of BIO101 on respiratory function. To accurately monitor changes in respiratory function throughout the course of treatment, whole-body plethysmography analysis (Emka Technologies, Paris, France) was performed on healthy control mice (C57Black10), untreated mdx mice, and mdx mice treated with BIO101.

[0065] The advantages of this technique are that it allows monitoring of awake animals that move freely within an airtight enclosure, and that it is performed non-invasively. As a result, stress on the animals from handling is reduced, and measurements can be repeated over a long period. Therefore, barometric plethysmography is frequently used to measure bronchial reactivity in small animals (Chong et al., 1998; Djuric et al., 1998; Hoffman et al., 1999).

[0066] Pressure fluctuations measured relative to a reference chamber allow us to define various respiratory parameters, such as the peak and timing of inspiratory and expiratory pressures, and a dimensionless quantity called Penh (enhanced Pause), which enables the evaluation of bronchoconstriction. More specifically, Penh, calculated from the chamber pressure signal (Pb), is an important indicator to acquire. This is because Penh fluctuations change in parallel with fluctuations in respiratory resistance, and therefore represent a predictive parameter regarding changes in the resistance characteristics of the respiratory system (Hamelmann et al., 1997; Bergren, 2001; Onclinx et al., 2003). The following values ​​were calculated from filtered Pb: maximum change in Pb during exhalation (PEP: peak expiratory pressure); maximum change in Pb during inspiration (PIP: peak inspiratory pressure); and time interval (TR). The Penh value was then calculated as follows: (PIP / PEP) × Pause Here Pause = (TE - TR) / TE Therefore, TE is the expiratory time (Adler et al., 2004).

[0067] Furthermore, peak inspiratory flow (PIF), peak expiratory flow (PEF), relaxation time (RT), and respiratory rate (BF) were also measured and illustrated.

[0068] Penh values ​​were measured before the start of treatment (D0), 30 days after the start of treatment (D30), and 60 days after the start of treatment (D60).

[0069] First, prior to treatment, the entire cohort of mdx mice (n=24) was compared to healthy mice C57Black10 (n=12). Awake mice were exposed to increasing doses of methacholine aerosol produced by a nebulizer, containing 0–40 mg / mL of methacholine in PBS. As expected and as previously demonstrated, mdx mice exhibited respiratory dysfunction (Huang et al., 2011; Gosselin et al., 2003; Gayraud et al., 2007; Ishizaki et al., 2008), particularly before or after administration of bronchoconstrictors, accompanied by a significant increase in methacholine-responsive Penh (p<0.001 and p<0.0001) (Figure 1A), associated with significantly lower peak inspiratory and peak expiratory flows (p<0.05 and p<0.001) (Figure 1B, 1C) compared to healthy mice. The relaxation time (RT) was also significantly longer in mdx mice compared to control mice (p<0.001 and p<0.0001) (Figure 1D). Inspiratory time (TI) and expiratory time (TE) values ​​remained comparable between these groups at each measurement time point (data not shown). The respiratory rate of mdx mice was also reduced compared to healthy mice (p<0.05 and p<0.0001) (Figure 1E).

[0070] Next, the mdx mice were divided into two groups: mdx mice that received no treatment (white squares); and mdx mice that would be administered the molecule BIO101 (black squares). As shown in Figures 2A, 2B, 2C, 2D, and 2E, these two groups exhibited identical capabilities in terms of respiratory function, with no significant differences among the various respiratory parameters measured. This indicates that, prior to treatment, all untreated mdx mice had the same respiratory profile.

[0071] Thirty days after the start of treatment, mdx mice treated with BIO101 showed a significant decrease in Penh (p<0.001) compared to mdx mice at D0, and the Penh curve in response to metacholine was comparable to that of healthy mice C57Black10 (Figure 3A). The decrease in Penh was associated with an increase in peak inspiratory flow, particularly at steady state or at low metacholine doses (p<0.05) (Figure 3C). Furthermore, a clear improvement in relaxation time was observed in mdx mice treated with BIO101 compared to untreated mdx mice (p<0.05) (Figure 3D). In contrast, peak inspiratory flow and respiratory rate remained unchanged compared to those measured at D0 (Figures 3B, 3E).

[0072] Sixty days after the start of treatment, BIO101 maintained its beneficial effect on respiratory function in mdx mice. Specifically, the effect observed 30 days after the start of treatment was subsequently confirmed. Mdx mice treated with BIO101 showed a significant decrease in Penh (p<0.0001) compared to untreated mdx mice, and the profile of Penh variability was similar to that of the control, regardless of the methacholine dose (Figure 4A). A significant improvement in peak inspiratory flow (p<0.05) was observed, along with a trend toward improvement in relaxation time (Figures 4B, 4D). In contrast, peak expiratory flow and respiratory rate remained unchanged (Figures 4B, 4E).

[0073] To longitudinally evaluate respiratory dysfunction in mdx mice, we compared Penh variability between D0 and D60 in healthy control mice, and between D0 and D60 in untreated mdx mice. As expected, Penh did not change between D0 and D60 in control mice (p=ns) (Figure 5A). In contrast, respiratory function was significantly worse in mdx mice at D60 compared to D0, as indicated by the increase in Penh in response to methacholine (p<0.0001) (Figure 5B).

[0074] At a methacholine dose of 40 mg / mL, the mean Penh values ​​in healthy control mice (n=12) were significantly lower than those observed in untreated mdx mice (n=23) (Penh = 0.72 and 1.42, respectively; p<0.0001) (Figure 6A). At D60, these Penh values ​​were higher in untreated mdx mice (n=12) compared to healthy control mice (n=12), with values ​​of 1.04 and 4, respectively (p<0.0001) (Figure 6B). Interestingly, treatment with BIO101 significantly reduced these values ​​compared to untreated mdx mice, resulting in a Penh value of 1.87 (p<0.0001) (Figure 6B). This treatment reduces Penh levels in mdx mice to a level that is not significantly different from that of healthy mice (Penh levels in mdx mice are 1.87 compared to 1.04 in C57Black10 mice; p<0.05).

[0075] b. Analysis of the effects of BIO101 on lung resistance, compliance, and elastance using a piston ventilator. To more directly determine the mechanical parameters of the respiratory system and the effects of two months of regular treatment with BIO101 (D60) on these parameters, dynamic lung resistance was measured in response to increasing methacholine doses using the piston ventilator system described above.

[0076] Mice were anesthetized and connected to a piston ventilator system via an endotracheal cannula. After initiating mechanical ventilation, each mouse was intraperitoneally injected with 0.1 mL of 10 mg / mL rocuronium bromide solution. The animals were ventilated at a respiratory rate of 150 breaths / min with a tidal volume of 10 mL / kg relative to a positive end-tidal pressure of 3 cmH2O. The respiratory mechanism was evaluated using a 1.2-second forced oscillation operation (2.5 Hz) and a 3-second broadband forced oscillation operation including 13 fundamental frequencies from 1 to 20.5 Hz. Respiratory system resistance (R) was calculated using a microcomputer-type central processing unit. This central processing unit was connected to the piston ventilator system and included software specifically for performing the above calculations. The two operations were performed alternately every 15 seconds after each spray of methacholine aerosol, and the change in response time of methacholine-induced bronchoconstriction was measured. In this way, lung resistance, compliance, and elastance could be determined.

[0077] Consistent with plethysmography results, untreated mdx mice were observed to have higher airway resistance than healthy C57Black10 mice (p<0.001) (Figure 7A). Sixty days after the start of treatment, as the methacholine dose increased from 0 to 20 mg / mL, mdx mice treated with BIO101 showed significantly lower airway resistance compared to untreated mdx mice (p<0.0001), which is comparable to the level of lung resistance observed in healthy control mice (Figure 8A). Similarly, regular treatment with BIO101 for two months significantly improved two parameters altered in mdx mice: lung compliance (p<0.0001) and elastance (p<0.0001) (Figures 7B, 7C). Specifically, at the highest methacholine dose tested (20 mg / mL), mdx mice exhibited significantly lower lung compliance compared to control mice C57Black10 (0.017 mL / cmH2O and 0.031 mL / cmH2O, respectively; p<0.0001). This deficiency in lung compliance was significantly corrected in mdx mice by treatment with BIO101 (0.029 mL / cmH2O; p<0.0001) (Figure 8B). Similarly, lung elastance was significantly lower in mdx mice compared to control mice C57Black10 (62.8 cmH2O / mL and 34.8 cmH2O / mL; p<0.05), and was maintained at levels comparable to healthy control mice and significantly higher than untreated mdx mice with treatment with BIO101 (34.8 cmH2O / mL and 69.1 cmH2O / mL; p<0.001) (Figure 8C).

[0078] 3. Conclusion These results demonstrate that treatment with BIO101 (50 mg / kg / day in drinking water) improves and sustains respiratory function in mdx mice (an animal model of Duchenne muscular dystrophy). This effect on respiratory function is related not only to respiratory parameters (duration and frequency of inspiration and expiration), as demonstrated by plethysmography results, particularly Penh measurements, but also to improvements in airway structure, as demonstrated in experiments using a piston ventilator system. Indeed, data from this system significantly demonstrate the beneficial effects of BIO101 treatment on mechanical respiratory parameters such as lung resistance, compliance, and elastance in mdx mice. These observations result in the protection of lung function from deterioration over time by treatment with plant ecdysones, particularly BIO101, in a mouse model of neuromuscular disease.

[0079] More generally, it should be noted that the embodiments for implementation and carrying out the present invention discussed above are described as non-limiting examples, and therefore other modifications are possible.

[0080] References Adler A, Cieslewicz G, Irvin CG. Unrestrained plethysmography is an unreliable measure of airway responsiveness in BALB / c and C57BL / 6 mice. J. Appl. Physiol. (1985). (2004); 97(1):286-92. Baydur A, Gilgoff I, Prentice W, Carlson M, Fischer DA. Decline in respiratory function and experience with long-term assisted ventilation in advanced Duchenne's muscular dystrophy. Chest (1990); 97:884-9. Benditt JO and Boitano LJ. Pulmonary Issues in Patients with Chronic Neuromuscular Disease, Am. J. Respir. Crit. Care Med. (2013); 187 (10):1046-55. Bergren DR. Chronic tobacco smoke exposure increases airway sensitivity to capsaicin in awake guinea pigs. J. Appl. Physiol. (2001); 90:695-704. Birnkrant DJ, Panitch HB, Benditt JO, Boitano LJ, Carter ER, et al. American College of Chest Physicians consensus statement on the respiratory and related management of patients with Duchenne muscular dystrophy undergoing anesthesia or sedation. Chest (2007); 132:1977-86. Chong BT, Agrawal DK, Romero FA, Townley RG. Measurement of bronchoconstriction using whole-body plethysmograph: comparison of freely moving versus restrained guinea pigs. J. Pharmacol. Toxicol. Methods (1998); 39:163-8. Djuric VJ, Cox G, Overstreet DH, Smith l, Dragomir A, Steiner M. Genetically transmitted cholinergic hyperresponsiveness predisposes to experimental asthma. Brain Behav. Immun. (1998); 12:272-84. Finder JD, Birnkrant D, Carl J, Farber HJ, Gozal D, et al. American Thoracic Society. Respiratory care of the patient with Duchenne muscular dystrophy: ATS consensus statement. Am. J. Respir. Crit. Care Med. (2004); 170:456-65. Gayraud J, Matecki S, Hnia K, Mornet D, Prefaut C, et al. Ventilation during airbreathing and in response to hypercapnia in 5 and 16 month-old mdx and C57 mice. J. Muscle Res. Cell Motil. (2007); 28(1):29-37. Gosselin LE, Barkley JE, Spencer MJ, McCormick KM, Farkas GA. Ventilatory dysfunction in mdx mice: impact of tumor necrosis factor-alpha deletion. Muscle Nerve (2003); 28:336-43. Hamelmann E, Schwarze J, Takeda K, Oshiba A, Larsen GL, Irvin CG, Gelfand EW. Noninvasive measurement of airway responsiveness in allergic mice using barometric plethysmography. Am. J. Respir. Crit. Care Med. (1997); 156:766-7 Hoffman AM, Dhupa N, Cimetti L. Airway reactivity measured by barometric whole-body plethysmography in healthy cats. Am. J. Vet. Res., (1999); 60:1487–9 Huang P, Cheng G, Lu H, Aronica M, Ransohoff RM, Zhou L. Impaired respiratory function in mdx and mdx / utrn(+ / -) mice. Muscle Nerve (2011); 43(2):263-7. [ PubMed ] [ Cross Ref ] Inkley SR, Oldenburg FC, Vignos PJ. Pulmonary function in Duchenne muscular dystrophy related to stage of disease. Am. J. Med. (1974); 56:297-3 Ishizaki M, Suga T, Kimura E, Shiota T, Kawano R, et al. Mdx respiratory impairment following fibrosis of the diaphragm. Neuromuscular. Disorder. (2008); 18(4):342-8 Lo Mauro A and Aliverti A. Physiology of respiratory disturbances in muscular dystrophies. Breathe (2016); 12(4):318-27. Matecki S, Rivier F, Hugon G, Koechlin C, et al. The effect of respiratory muscle training with CO2breathing on cellular adaptation of mdx mouse diaphragm. Neuromuscul. Disord. (2005); 15:427-36. Mayer OH, Finkel RS, Rummey C, Benton MJ, Glanzman AM, et al. Characterization of Pulmonary Function in Duchenne Muscular Dystrophy, Pediatr. Pulmonol. (2015); 50:487-94. McKim DA, Road J, Avendano M, Abdool S, Cote F, et al. Canadian Thoracic Society Home Mechanical Ventilation Committee. Home mechanical ventilation: a Canadian Thoracic Society clinical practice guideline. Can. Respir. J. (2011); 18:197-215. Miller RG, Jackson CE, Kasarskis EJ, England JD, Forshew D, et al. Quality Standards Subcommittee of the American Academy of Neurology. Practice parameter update: the care of the patient with amyotrophic lateral sclerosis: drug, nutritional, and respiratory therapies (an evidence-based review): report of the Quality Standards Subcommittee of the American Academy of Neurology. Neurology (2009); 73:1218-26. Mosqueira M, Baby SM, Lahiri S, Khurana TS. Ventilatory chemosensory drive is blunted in the mdx mouse model of Duchenne Muscular Dystrophy (DMD). PLoS One. (2013); 8(7):e69567 Onclinx C. Relationship between total pulmonary resistance and Penh value as a function of the anatomical location of the airway obstruction (in-depth study diploma thesis). University of Liège, Faculty of Veterinary Medicine: Liège, (2003), 24p. Smith PE, Calverley PM, Edwards RH, Evans GA, Campbell EJ. Practical problems in the respiratory care of patients with muscular dystrophy. N.Engl. J.Med. (1987); 316:1197-205. Vianello A, Bevilacqua M, Salvador V, Cardaioli C, Vincenti E. Long-term nasal intermittent positive pressure ventilation in advanced Duchenne's muscular dystrophy. Chest (1994); 105:445-8.

Claims

1. A composition comprising at least one phytoecdysone and / or at least one semi-synthetic derivative of phytoecdysone for its use in a mammal in the treatment of respiratory dysfunction resulting from an acquired or inherited neuromuscular disease or respiratory dysfunction associated with bronchial hyperresponsiveness.

2. 2. A composition for use thereof according to claim 1, comprising 20-hydroxyecdysone and / or at least one semi-synthetic derivative of 20-hydroxyecdysone.

3. 20-hydroxyecdysone in the form of a plant extract or an extract of a plant part, the plant is selected from plants containing at least 0.5% 20-hydroxyecdysone by dry weight of the plant; 3. A composition for use thereof according to claim 2, wherein the extract contains at least 95%, preferably at least 97%, of 20-hydroxyecdysone.

4. Notably, the composition for use thereof according to claim 3 contains 0-0.05% of the dry weight of the extract of impurities that may affect the safety, availability or efficacy of the extract for pharmaceutical application.

5. 5. The composition for its use according to claim 3 or 4, wherein the plant is selected from Stemmacantha carthamoides, Cyanotis arachnoidea and Cyanotis vaga.

6. The composition for use thereof according to any one of claims 1 to 5, wherein the respiratory dysfunction is due to a neuromuscular disease of the motor neurons and / or of the neuromuscular junction and / or of the striated muscles.

7. The composition for use according to any one of claims 1 to 6, wherein the respiratory dysfunction is associated with disorders of striated and / or smooth muscles.

8. The composition for use thereof according to any one of claims 1 to 5, wherein said bronchial hyperresponsiveness is related to the function of bronchial smooth muscle.

9. 9. The composition for use thereof according to any one of claims 1 to 8, wherein the respiratory dysfunction is related to the state of at least one of the respiratory parameters selected from Penh value, peak inspiratory flow, peak expiratory flow, relaxation time, and respiratory rate.

10. The composition for use thereof according to any one of claims 1 to 9, wherein said respiratory dysfunction is related to the state of at least one of the mechanical parameters of the lung tissue.

11. 11. The composition for use according to claim 10, wherein the respiratory dysfunction is associated with decreased lung compliance and / or increased lung resistance and / or decreased lung elastance.

12. The composition for use thereof according to any one of claims 1 to 11, wherein the phytoecdysones are administered in a dose of 3 to 15 mg / kg / day in humans.

13. 13. The composition for use thereof according to any one of claims 1 to 12, wherein the phytoecdysones are administered in one or more divided doses of 200 to 1000 mg / day to an adult human and in one or more divided doses of 5 to 350 mg / day to a human child or infant.

14. General formula (I): 【Chemistry 1】 wherein: VU is a carbon-carbon single bond and Y is a hydroxyl group or hydrogen, or VU is an ethylenic C=C bond; X is oxygen; Q is a carbonyl group; R １ Is: (C １ -C ６ ) W (C １ -C ６ ) group; (C １ -C ６ ) W (C １ -C ６ ) W (C １ -C ６ ) group; (C １ -C ６ ) W (C １ -C ６ ) CO ２ (C １ -C ６ ) group; (C １ -C ６ ) A group, where A is OH, MeO, (C １ -C ６ ), N(C １ -C ６ ), CO ２ (C １ -C ６ represents a heterocycle optionally substituted with a group of the type CH ２ Br groups; The composition for use thereof according to any one of claims 1 to 13, wherein W is a heteroatom selected from N, O and S, preferably O, more preferably S.

15. In the general formula (I): Y is a hydroxyl group; R １ Is: (C １ -C ６ ) W (C １ -C ６ ) group; (C １ -C ６ ) W (C １ -C ６ ) W (C １ -C ６ ) group; (C １ -C ６ ) W (C １ -C ６ ) CO ２ (C １ -C ６ ) group; (C １ -C ６ ) A group, where A is OH, MeO, (C １ -C ６ ), N(C １ -C ６ ), CO ２ (C １ -C ６ ) type groups; 15. The composition for use thereof according to claim 14, wherein W is a heteroatom selected from N, O and S, preferably O, more preferably S.

16. Said at least one compound of said general formula (I) is: No. 1: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-10,13-dimethyl-17-(2-morpholinoacetyl)-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-6-one; No. 2: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-17-[2-(3-hydroxypyrrolidin-1-yl)acetyl]-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-6-one; No. 3: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-17-[2-(4-hydroxy-1-piperidyl)acetyl]-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-6-one; No. 4: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-17-[2-[4-(2-hydroxyethyl)-1-piperidyl]acetyl]-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-6-one; No. 5: (2S,3R,5R,10R,13R,14S,17S)-17-[2-(3-dimethylaminopropyl(methyl)amino)acetyl]-2,3,14-trihydroxy-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-6-one; No. 6: 2-[2-oxo-2-[(2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-10,13-dimethyl-6-oxo-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-17-yl]ethyl]ethylsulfanyl acetate; No. 7: (2S,3R,5R,10R,13R,14S,17S)-17-(2-ethylsulfanylacetyl)-2,3,14-trihydroxy-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-6-one; No. 8: (2S,3R,5R,10R,13R,14S,17S)-2,3,14-trihydroxy-17-[2-(2-hydroxyethylsulfanyl)acetyl]-10,13-dimethyl-2,3,4,5,9,11,12,15,16,17-decahydro-1H-cyclopenta[a]phenanthren-6-one 16. The composition for use thereof according to claim 14 or 15, selected from:

17. General formula (II): 【Chemistry 2】 A composition for use thereof according to any one of claims 1 to 16, comprising at least one compound of the formula:

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