Mucoadhesive polymers: Design of S-protected thiolated cyclodextrin-based hydrogels.

AIM: This study aims to design chemically crosslinked thiolated cyclodextrin-based hydrogels and to evaluate their mucoadhesive properties via mucosal residence time studies on porcine small intestinal mucosa and on porcine buccal mucosa. METHODS: Free thiol groups of heptakis(6-deoxy-6-thio)-ß-cyclodextrin (ß-CD-SH) were S-protected with 2-mercaptoethanesulfonic acid (MESNA) followed by crosslinking with citric acid. Cytotoxicity was assessed by hemolysis as well as resazurin assay. Hydrogels were characterized by their rheological and mucoadhesive properties. Ritonavir was employed as model drug for in vitro release studies from these hydrogels. RESULTS: The structure of S-protected ß-CD-SH was confirmed by IR and 1H NMR spectroscopy. Degree of thiolation was 390 ± 7 µmol/g. Hydrogels based on native ß-CD showed hemolysis of 12.5 ± 2.5 % and 13.6 ± 2.7 % within 1 and 3 h, whereas hemolysis of just 3.5 ± 2.8 % and 3.9 ± 3.0 % was observed for the S-protected thiolated CD hydrogels, respectively. Both native and S-protected thiolated hydrogels showed minor cytotoxicity on Caco-2 cells. Rheological investigations of S-protected thiolated ß-CD-based hydrogel (16.2 % m/v) showed an up to 13-fold increase in viscosity in contrast to the corresponding native ß-CD-based hydrogel. Mucosal residence time studies showed that thiolated ß-CD-based hydrogel is removed to a 16.6- and 2.4-fold lower extent from porcine small intestinal mucosa and porcine buccal mucosa in comparision to the native ß-CD-based hydrogel, respectively. Furthermore, a sustained release of ritonavir from S-protected thiolated ß-CD-based hydrogels was observed. CONCLUSION: Because of their comparatively high mucoadhesive and release-controlling properties, S-protected thiolated ß-CD-based hydrogels might be promising systems for mucosal drug delivery.

Intraoral Drug Delivery: Highly Thiolated κ-Carrageenan as Mucoadhesive Excipient.

AIM: This study aims to design a novel thiolated κ-carrageenan (κ-CA-SH) and evaluate its potential as an excipient for the design of mucoadhesive drug delivery systems. METHODS: Native κ-carrageenan (κ-CA) was thiolated with phosphorous pentasulfide in sulfolane and characterized via 1H NMR, FTIR, as well as Ellman's test. Cytotoxicity was assessed via resazurin assay. In vitro release of the model drug, benzydamine hydrochloride, was determined. Tensile and mucosal residence time studies were performed on buccal and small intestinal mucosa. Mucoadhesive features were investigated via rheological studies with freshly isolated porcine mucus. RESULTS: Thiolated κ-CA (κ-CA-SH) with 1213.88 ± 52 µmol/g thiol groups showed no cytotoxicity at a concentration of 1% (m/v) and low cytotoxicity up to 2% (m/v). Benzydamine hydrochloride showed slow release in solution for both polymers. Tensile studies on buccal and intestinal mucosa showed an up to 2.7-fold and 7.7-fold enhancement in the maximum detachment force (MDF) and total work of adhesion (TWA) of κ-CA-SH vs. κ-CA, respectively. The κ-CA-SH exhibited an up to 4.4-fold improved dynamic viscosity with mucus and significantly prolonged residence time on mucosa compared to native κ-CA. CONCLUSION: Since highly thiolated κ-CA shows a slow release of positively charged active pharmaceutical ingredients and enhanced mucoadhesive properties, it might be a promising excipient for local drug delivery in the oral cavity.

Betaine-modified hydroxyethyl cellulose (HEC): A biodegradable mucoadhesive polysaccharide exhibiting quaternary ammonium substructures.

The aim of this study was to improve the mucoadhesive properties of hydroxyethyl cellulose (HEC) via the covalent attachment of betaine. Synthesis was carried out through esterification of HEC utilizing N-chlorobetainyl chloride. Betaine-modified HEC was characterized via FTIR and NMR analyses, ester quantification and zeta potential measurements. Enzymatic degradation and cell viability were also investigated. Moreover, rheological and mucoadhesive properties were evaluated. FTIR and NMR analyses confirmed the covalent attachment of betaine to HEC. Betaine-modified HEC contained 228.45±11.63 µmol/g ester bonds and its zeta potential was 0.37±0.19 mV. Enzymatic degradation studies showed the ability of lipase to cleave off betaine from HEC. Cytotoxicity studies demonstrated that betaine-modified HEC is up to a concentration of 0.3% not toxic. In comparison to unmodified HEC, betaine-modified HEC showed with mucus a 2.3- and 4-fold higher viscosity within 3 h and 6 h, respectively. Furthermore, betaine-modified HEC exhibited 23.5-fold higher mucoadhesive properties on porcine intestinal mucosa compared to unmodified HEC. In conclusion, betaine-modified HEC might be a useful biodegradable mucoadhesive polymer.

Imine bond formation as a tool for incorporation of amikacin in self-emulsifying drug delivery systems (SEDDS).

AIM: The aim was to develop a self-emulsifying drug delivery system (SEDDS) for amikacin via imine bond formation with hydrophobic aldehydes. METHODS: Trans-2, cis-6-nonadienal, trans-cinnamaldehyde, citral and benzaldehyde were conjugated to amikacin at pH 8.5. Based on results of precipitation efficiency, Fourier-transform infrared spectroscopy (FTIR) and NMR analysis, amikacin-trans-cinnamaldehyde conjugates were further characterized regarding log Poctanol/water via HPLC. The release of amikacin from the amikacin-trans-cinnamaldehyde conjugates was examined through in vitro incubation with bovine serum albumin (BSA). SEDDS containing the amikacin-trans-cinnamaldehyde conjugates were tested regarding mean droplet size (MDS), polydispersity index (PDI), log DSEDDS/release medium and cell viability. RESULTS: Trans-cinnamaldehyde formed the most hydrophobic conjugates with amikacin whereas benzaldehyde did not form hydrophobic conjugates at all. Imine bond formation was confirmed by FTIR and NMR analysis. The highest increase in log P was achieved for the amikacin-trans-cinnamaldehyde conjugate in a molar ratio of 1:5, shifting from -8.58 up to 1.59. Incubation of this conjugate with BSA led to the formation of BSA-trans-cinnamaldehyde releasing in turn amikacin. SEDDS based on Capmul MCM, Cremophor EL and propylene glycol containing the conjugate demonstrated a MDS of 61.4 nm and PDI of 0.265. Log DSEDDS/release medium was calculated to be 3.38. Cell viability studies showed very good tolerability of conjugate loaded SEDDS in concentrations of 0.1% - 0.5%. CONCLUSION: Imine bond formation of amikacin with trans-cinnamaldehyde and the incorporation of the resulting conjugate into SEDDS represents a promising strategy for oral delivery of amikacin.

Imine bond formation: A novel concept to incorporate peptide drugs in self-emulsifying drug delivery systems (SEDDS).

HYPOTHESIS: Because of its hydrophilic character the peptide drug Polymyxin B (PMB) cannot be incorporated in lipophilic nanocarrier systems such as self-emulsifying drug delivery systems (SEDDS) for oral administration. Due to the formation of imine conjugates between the primary amino groups of PMB and the carbonyl group of cinnamaldehyde, however, drug lipophilicity might be sufficiently raised for incorporation in SEDDS. METHODS: Imine bonds were formed between the primary amino groups of PMB and the carbonyl group of cinnamaldehyde. PMB-cinnamaldehyde conjugate was characterized regarding degree of substitution, log P and release of PMB due to interaction with bovine serum albumin (BSA), SEDDS loading and cell viability. RESULTS: 87.1% of primary amines formed imines with cinnamaldehyde. Log P was increased 69.183 - folds. BSA triggered release of PMB was 45.2%, 64.9% and 80.6% within 16 h. Log DSEDDS/Release medium of PMB-cinnamaldehyde conjugate was 3.4. CONCLUSION: According to these findings, the concept of imine bond formation with cinnamaldehyde can be considered as a novel concept for increasing lipophilicity of the hydrophilic antibiotic peptide PMB.

Arginine-based cationic surfactants: Biodegradable auxiliary agents for the formation of hydrophobic ion pairs with hydrophilic macromolecular drugs.

WORKING HYPOTHESIS: It was the hypothesis of this study that esters of arginine (Arg) with medium and long chain aliphatic alcohols are biodegradable and less cytotoxic than well-established cationic surfactants being used for hydrophobic ion pairing (HIP) with hydrophilic macromolecular drugs. EXPERIMENTS: Arg was linked to nonan-1-ol and hexadecan-1-ol (C9 and C16) via an ester linkage. The newly formed Arg-nonyl ester (ANE) and Arg-hexadecanoyl ester (AHE) surfactants were evaluated regarding critical micelle concentration (CMC) using pyrene fluorescent method, cytotoxicity on human colorectal adenocarcinoma-derived cells (Caco-2) and biodegradability at the concentrations of 2.5 and 5 mg/mL using 2500 Nα-benzoyl-l-arginine ethyl ester hydrochloride (BAEE) units/mL of trypsin. Furthermore, in order to evaluate their potential for HIP, heparin and daptomycin were used as model polysaccharide and peptide drugs, respectively. FINDINGS: Chemical structures of ANE and AHE surfactants were confirmed by FTIR, 1H NMR, and LC-MS. CMC of ANE was 7.5 mM and CMC of AHE was 2 mM. Arg-surfactants were not cytotoxic below their CMC. At CMC and above CMC, ANE was significantly (P < 0.05) more cytotoxic than AHE. ANE in both concentrations was degraded ˃98% within 48 h. The degradation of AHE at lower concentration was ˃97% and about 50% at higher concentration. Arg-surfactants were able to efficiently precipitate heparin and daptomycin from corresponding aqueous solutions. CONCLUSION: Arg-surfactants being biodegradable and less toxic seems to be a promising alternative to well-established cationic surfactants for the formation of hydrophobic ion pairs (HIPs) with hydrophilic macromolecular drugs.

Improved Intestinal Mucus Permeation of Vancomycin via Incorporation Into Nanocarrier Containing Papain-Palmitate.

The aim of this study was to improve intestinal mucus permeation of a peptide antibiotic via incorporation into papain-palmitate-modified self-emulsifying drug delivery systems (SEDDS) as nanocarrier. Vancomycin as a peptide antibiotic was lipidized by hydrophobic ion pair formation using sodium bis-2-ethylhexyl-sulphosuccinate before incorporation in SEDDS comprising Capmul MCM, propylenglycol, and Kolliphor EL (2:1:2). As mucolytic agent, 0.5% papain-palmitate was introduced in SEDDS formulation containing the vancomycin-sodium bis-2-ethylhexyl-sulphosuccinate ion pair. The formulation was evaluated regarding droplet size, zeta potential, and cytotoxicity using Caco-2 cells previous to intestinal mucus permeation studies using Transwell diffusion and rotating tube method. The hydrophobic ion pair product yielded from surfactant to drug ratio of 3:1 provided a 25-fold increase in lipophilicity, drug payload in SEDDS of 5%, and log DSEDDS/release medium of 2.2. The formulation exhibited a droplet size and zeta potential of 221.5 ± 14.8 nm and -4.2 ± 0.8 mV, respectively. Cytotoxicity study showed that SEDDS formulations were not toxic. Introducing 0.5% papain-palmitate increased the mucus permeability of SEDDS 2.8-fold and 3.3-fold in Transwell diffusion and rotating tube studies, respectively. According to these results, papain decorated SEDDS might be a potential strategy to improve the mucus permeating properties of peptide antibiotics.

Self-emulsifying drug delivery systems: Impact of stability of hydrophobic ion pairs on drug release.

The aim of this study was to evaluate the impact of stability of hydrophobic ion pairs (HIPs) in gastrointestinal (GI) fluids on their release from self-emulsifying drug delivery systems (SEDDS). HIPs of leuprolide (LEU), insulin (INS) and bovine serum albumin (BSA) were formed using various mono- and di-carboxylate surfactants i.e. sodium deoxycholate (SDC), sodium dodecanoate (SDD), sodium stearoyl glutamate (SSG) and pamoic acid di-sodium salt (PAM). HIPs were evaluated regarding precipitation efficiency, log Pn-butanol/water and dissociation behavior at various pH and ionic strength. Solubility studies of these HIPs were accomplished to identify suitable solvents for the formulation of SEDDS. Subsequently, HIPs were incorporated into SEDDS followed by characterization regarding zeta potential, stability and log DSEDDS/release medium. Independent from the type of (poly)peptides, PAM showed most efficient HIP properties among tested surfactants. The highest encapsulation efficiency with PAM was achieved at molar ratios of 1:1 for LEU, 1:3 for INS and 1:50 for BSA and log Pn-butanol/water of HIPs were increased at least 2.5 units. Dissociation studies showed that LEU-PAM, INS-PAM, BSA-PAM complexes were dissociated within 6 h up to 25%, 60% and 85% in GI fluids, respectively. These HIPs were successfully incorporated into SEDDS exhibiting negative zeta potential and high stability for 4 h. Log DSEDDS/release medium of LEU-PAM, INS-PAM, BSA-PAM complexes were 2.4 ±â€¯0.7, 2.1 ±â€¯0.62 and 1.6 ±â€¯0.45, respectively. Findings of this study showed that stability of HIPs has great impact on log DSEDDS/release medium and consequently on their release from SEDDS.

Trypsin decorated self-emulsifying drug delivery systems (SEDDS): Key to enhanced mucus permeation.

It was the aim of this study to prepare trypsin decorated mucus permeating self-emulsifying drug delivery systems (SEDDS). Lipophilicity of enzyme was increased by hydrophobic ion pairing (HIP) with the anionic surfactants sodium dodecyl sulfate (SDS), sodium taurocholate (ST) and sodium deoxycholate (SDO) to facilitate its incorporation in SEDDS. Blank SEDDS and trypsin decorated SEDDS were characterized regarding droplet size, polydispersity index (PI), zeta potential and proteolytic activity using Nα-benzoyl-l-arginine ethyl ester (BAEE) assay. Log DSEDDS/release medium of each complex was determined to assess its affinity towards SEDDS oily droplet upon emulsification. Ability of trypsin decorated SEDDS to enhance mucus permeation was studied on mucus gel from porcine small intestine for the period of 4 h at 37 °C. Degree of enzyme precipitation via HIP was 94.5%, 85.7% and 48.2% for SDS, ST and SDO complex, respectively. SEDDS composed of 50% (w/w) cremophor EL, 20% (w/w) captex 300, and 30% (w/w) propylene glycol with a complex payload of 1% (w/w) exhibited a droplet size in the range of 29.92 ±â€¯0.09 nm to 39.15 ±â€¯0.37 nm, a polydispersity index of 0.116-0.265 and zeta potential in the range of -2.36 mv to -4.25 mv. The enzymatic activity of trypsin complexed with SDO, SDS and ST in SEDDS was 51.9%, 44.8%, and 40.7% respectively, of the corresponding activity of free trypsin. Log D values of trypsin, SDS, ST and SDO complex were -2.73, 1.97, 1.89 and 1.68, respectively, suggesting higher lipophilicity of trypsin complexes as compare to free trypsin and ability to reside on SEDDS droplets. Enzyme decorated SEDDS improved mucus permeation 1.6- to 2.6-fold in comparison to blank SEDDS. Results demonstrated that decorating SEDDS with trypsin can be a promising technique to improve their mucus permeating properties.

PD-L1 Status in Refractory Lymphomas.

Targeted immunotherapy based on PD-1/PD-L1 suppression has revolutionized the treatment of various solid tumors. A remarkable improvement has also been observed in the treatment of patients with refractory/relapsing classical Hodgkin lymphoma (cHL). We investigated PD-L1 status in a variety of treatment resistant lymphomas. Tumor samples from 78 patients with therapy resistant lymphomas were immunohistochemically (IHC) investigated for the expression of PD-L1 using two antibody clones (SP142 and SP263, Ventana). Thirteen PD-L1+ cases were further analyzed for gene copy number variations (CNV) by NGS and for PD-L1/JAK2/PD-L2 co-amplification using fluorescent in-situ hybridization assay (FISH). PD-L1 positivity (≥5% positive cancer cells, IHC) was present in 32/77 (42%) and 33/71 cases (46%) using SP142 and SP263 antibodies, respectively. Concordance between the two anti-PD-L1 clones was high with only three (4%) discrepant cases. The strongest and consistent (10/11 cases) expression was observed in cHL and primary mediastinal B-cell lymphomas (3/3). Diffuse large B-cell lymphomas (DLBCL) were frequently positive (13/26) irrespective of subtype. Follicular (1/8), peripheral T-cell (3/11) and mantle cell (1/8) lymphomas were rarely positive, while small lymphocytic lymphoma/CLL and marginal zone lymphomas were consistently negative (3/3). Co-amplification/CNVs of PD-L1/JAK2/PD-L2 were observed in 3 cases of DLBCL and cHL, respectively. Of note, all three cHL-amplified cases were positive by FISH, but not by NGS. Since only a fraction of the IHC positive lymphoma cases were positive by FISH and NGS assays, other mechanisms are involved in PD-L1 upregulation, especially in DLBCL. FISH assay may be more suitable than NGS assay for determination of PD-L1 alterations in cHL.

[Treatment of Pseudomonas aeruginosa in cystic fibrosis in a child]. / Tretman pseudomonas aeruginosa kod oboljele djece sa cisticnom fibrozom.

The report deals with the case of a 10-year-old girl with chronic cystic fibrosis. She has been repeatedly treated at the hospital. She has been hospitalized due to respiratory deterioration. Cystic fibrosis is a rare disease, inherited autosomaly recessively, but is very complex in terms of diagnostic and treatment. Fibrosis is the formation of scar tissue due to injury or long term inflammation. The diagnosis is confirmed based on a clinical picture of the child, measure of Chloride in the sweat, chest X-ray, CT thorax, laboratory findings--genetic confirmation CFTR genes. The diagnosis is originally set when she was 4 years old. She is now admitted due to a deterioration of the main disease. Five days before the admission, the girl had a higher bodily temperature, cough and difficult breathing. Due to the deteriorated general condition and the respiratory insufficiency and respiratory acidosis in blood gas analysis, the girl was intubated and put on the complete mechanical ventilation (IPPV). Since the girl is a chronic patient with bronchiectasie chronic walls of bronchi changes full of the mucus, who is not responding to conservative treatment (antibiotics), therapeutic and diagnostic flexible bronchoscopy had to be performed, resulting in a gram-negative bacteri Pseudomonas aeruginosa--a typical bacteri for chronically sick C. F. patient. Pseudomonas aeruginosa is typically acquired in early childhood. This bacteria is giving progressive lung disease and often aggravates morbidity and mortality. So the main thing as a respiratory management is prevention of lung infection with this bacteria. A Pseudomonas therapy was prescribed according to the sensitive antibiogram, (Garamycin). Antibiotics are crucial to treating cystic fibrosis lung infections. Therapy with an amynoglicoside in combination with a B-lactam or a quinolone antibiotic is standard. It is a difficult to deliver a high doses at these antibiotics via the iv. route without significant systemic adverse events (otoxicity and nephrotoxicity). A reformulation of the aminoglycoside antibiotic tobramicin or garamycin therapy is solution for inhalation. To be well established infections the suppression of Pseudomonas aeruginosa has been shown to lead to decreasing same bacteria and benefits lung function from antibiotic therapy in a way that can be maintained over extended period. During bronchoscopy was given locally on changes mucous pulmozyme (to destroy a very hard mucous) and garamycin. So, after taking out a lot of mucus, it was later continued spontaneously. Control chest x ray and blood gas analysis are now very improved.

[Sarcoma Ewing left hemithorax]. / Sarcoma Ewing lijeve strane grudnog kosa.

A case of sarcoma Ewing hemithorax lst sin has been reported in an 8-year-old girl. The girl was admitted in our hospital with referral diagnosis Pleuropneumonia exudativa left side. The girl was sent from Nova Bila with the foregoing diagnosis. The difficulties started in early April this year with high body temperature, pains in the left side of the chest. After a detailed anamnesis, clinical picture of the girl and radiological examinations/ chest x ray, CT scan of the chest and operative findings/ sarcoma Ewing diagnosis was confirmed. The disease is very rare, while diagnostics and course are complex. The girl was admitted at the Pulmology Department of the Pediatric Clinic on 16.03.2005. At the Department, the girl was medically treated with three antibiotics: cephalosporins of third generation, ceftriaxon (Longacef), aminoglycoside Amikacin and antistaphyloccoc therapy (Orbenin). Due to a dull sound on percussion and weaker auscultatory breathing on the left, and radiological shading of the left chest, a CT of the thorax was performed. It showed expansive changes of the left chest. Also performed was a pleural punction of the left chest, after which around 500 ml sero-hemorrhagic content was taken out and sent for patohistology analysis. After a pre-operative preparation, the operation was performed on 28.03.2005, which is when a fat-like tumorous mass was found completely filling up the pleural cavum and by its weight exerting pressure on the lungs. In the projection of VI and VII rib, the described mass infiltrates the thoracic wall. The tumor extirpation "in toto" was performed and sent for patohistology analysis, ex tempore. Also performed was a pleuropectomy in the projection of the wall infiltration, as well as a partial resection of the VII rib. Also performed was a decortication of the visceral pleurae. The post-operative course has been passing normally and with good pleuropulmonary findings, with the wound healing per primum. Patohistology results: sarcoma Ewing hemithorax lat. sin. The girl continued her cytostatic therapy in Zagreb following the operation.

[Pulmonary abscess]. / Apscesi pluca.

A case of lung abscessi has been reported in 10 years old child, boy. This lung disease is uncommon, but treatment is complex. A lung abscess is a suppurative process resulting in destruction of the pulmonary parenchyma and formation of a cavity containing purulent material. The child was already treated in the hospital in Bihac since 19.07.2004. to 04.08.2004. Lung abscess was secondary caused by staphylococcus, started as panaritium second finger. The diagnosis is generally made by roentgenographic examination when a cavity with a fluid level surrounded by alveolar infiltration is demonstrated. After a few consultations with thoracal surgeon conservative treatment was continued Vankomicin 40 days and Funzol, later Stanicid 10 days. Brronchosacopy to faciliate drainage or to obtain culture is controversial so the same wasn't done. Surgical drainage of a lung abscess is almost never indicated and resection should be considered only in a children with recurrent hemoptysis, repeated episodis of infection. Serial chest roentgenograms show gradual diminution of the abscessus over a period of several weeks during hospitalization. Last one chest X ray shows as sequely air cyst on the left side. X ray of the second finger shows osteitis of the second phalange. After 40 days the child was discharged with recommendation for follow up by thoracal surgeon next 6 months.

[Congenital lobar emphysema]. / Kongenitalni lobarni emfizem.

BACKGROUND: Congenital lobar emphysema is characterized by overinflation of pulmonary lobe and is caused by localised bronchial obstruction. The disease may result in severe respiratory distress in early infancy. METHODS: Records of children, with congenital lobar emphysema, who were treated at our department between 1997-2003, were reviewed. RESULTS: Three males, aged 16 days till 1 year, were diagnosticed as congenital lobar emphysema. Presenting symptoms were dyspnea in 2 patients, cyanosis in 2 patients, wheesing in 2 patients, recurrent respiratory tract infection in 2 patients. Chest x rays and computerized tomography scans showed hyperinflation of the affected lobe in all patients. The affected sites were left upper lobe in 2 patients and right upper lobe in 1 patient. All patients underwent lobectomy. CONCLUSION: It is necessary to suspect on this uncommon anomaly in early infancy. The diagnosis was established by chest x rays and CT scans. Intensive therapy and urgent lobectomy, in severe respiratory distress, are condition for successfully treatment.

[Unusual therapeutic approach in treatment of pulmonary tuberculosis in six year old girl]. / Nuobicajen terapljski pristup lijecenja plucne forme tuberkuloze kod sestogodisnje djevojcice.

We report about interesting case of treatment of pulmonary tuberculosis in 6-year old girl. Antituberculotic therapy induced toxic hepatitis (rise of transaminases, indirect hiperbilirubinemia). She was treated with streptomycin, etambutol, ciprofloxacyn and systemic corticosteroides. Two gallbladder stones and high-density sediment were diagnosed after ultrasound examination which could be a side effect of the therapy or previous findings.

[Treatment of myeloproliferative diseases]. / Tretman mijeloproliferativnih bolesti.

The modern treatment of the myeloproliferative disease is based on knowledge about pathogenic tumour's growing up and basically the treatment is based on the use of cytoxic therapy. The mix of cytotoxic is bringing up to the reducing of disease at the some percentages. Related the above disease in practice interferons showed themselves quite efficiently and in practice of allogenic transplantation of marrow bone is gave and a possibility of recovery of the chronic myeloid leukemia. The use of alias "the smart medicines" are made a blockade of transgenes signals and inhibiting induced pathologic coagulation of enzymes and responsible for a cell homeostasis (cell's growth) opened a new perspective in the treatment of malignant haematological diseases. The right direction of treatment' strategy for sure is bringing to the survival extension.