|Results:The efficacy of the drug in the treatment of the patients are evaluated as comparison of the following values before and after the treatment:
a) Serum bilirubin, find that the mean value of serum bilirubin level of the patients before treatment is 1.67 mg % (normally 0.2 – 1mg%), with after treatment the mean value serum bilirubin of the patient showed highly significant lower to the normal range 0.83 mg % (p value < 0.001 which is highly signification).
b)Serum liver enzymes (indicators of liver cells insult):
1-Laboratory analysis of :
ALT (Alanine transaminase) find that the means value of serum ALT level of the patients before treatment is 91.13 units / ml (normally is up to 40units / ml), while after treatment the mean value of serum ALT level is 26.56 units / ml which shows highly significant lowering to the normal range (P value < 0.001 which is highly significant).
AST (Aspartate transaminase) finding that the mean value of serum AST level of the patients before treatment is 108.68 units / ml (normally is up to 37 units / ml), while after treatment the mean value of serum AST level is 36.76 units / ml which shows highly significant lowering to the normal range (P value < 0.001 which is highly significant).
2- Ultrasonography examination (U/ S)
a) Ultrasongraphic examination of liver condition (size & parenchymal affection) Find that, U / S liver before the treatment (73% of patients present with mild liver enlargement, 21.5% with moderate liver enlargement & 7% with shrunken cirrhotic liver), while after the treatment shows significant improvement of liver condition as 62.5% of patients present normal U / S liver findings, 29% of patients show just mild enlargement, only 1.5% of shrunken liver & 1% present moderately enlargement)
N.B the rest of shrunken liver patients most probably is missing cases.
b) Ultrasonography examination of portal vein diameter (normally : 8 – 12 mm).
Find that, U/S portal vein the mean value of diameter before the treatment is 13.38mm, while after treatment shows highly significant improvement & normalization of portal vein diameter & pressure as the mean value of diameter became 10.31 mm (normal diameter) (P value < 0.001 which is highly significant).
c) Ultrasonography examination of spleen (size), find that, U/S spleen size before the treatment (58% of patients present with moderate enlargement, 28% of patients show mild enlargement, 4% of patients with marked spleen enlargement & 7% of patients have normal spleen size), while after the treatment, the patients show significant improvement of spleen size (51% of patients show mild enlargement, 45% of patients have normal spleen size & only 1% with moderate spleen enlargement).
D) Ultrasonography examination for ascites, find that, U/S ascites examination before the treatment (2% of patients with minimal ascites & 1% of cases show moderate ascites 1.5% of patients present tense ascites), while after the treatment, the patients show significant improvement of ascites (only 1.5% of patients show minimal ascites).
3- HCV PCR (Polymerase Chain Reaction) Quantitative Assessment
Three monthly HCV PCR Quantitative assessment of blood HCV viraemia of the patients find that complete remission (CR) of HCV viraemia in 101 patients (50.5% by giving negative HCV PCR (Confirmed by repeating the negative PCR, three times successively, two weeks apart between each negative result), the cured CR patients are 101 cases
(50.5%) are classified into four groups according to the duration of treatment given, find that. (Most of the patients, 56 cases (28%) are cured in duration of 1-1.5 years of treatment, 35 cases (17.5%) are cured in duration of 1 year of treatment, 7 cases (3.5%) are cured in duration more than 1.5 years & minimal cases, 3 (1.5%) are cured in duration less than 1 year), the efficacy of the drug treatment & its cure duration depends on the level of HCV viraemia at the beginning of treatment. There are partial remission (PR) improvement of HCV viraemia in 99 cases (49.5%) showing significant improvement of HCV PCR levels most of them 48 cases (24%) are improved in duration of less than 1 year, 47 cases (23.5%) are improved in duration of 1 year, 2 cases (1%) are improved in duration of 1-1 ½ year and
2 cases (1%) are improved in duration more than 1.5 year), there results depend on the level of HCV viraemia at the start of treatment (Figure 1).
Relation between PCR – HCV viraemia at the beginning of treatment and duration of CR or PR are shown in (Figure 2).
4- Liver Biopsy & Histopathological examination:
a) HCV activity scoring system (normal score is 0/9) HCV activity before the treatment, find that most of the patients, 131 cases (65.5%) have high HCV activity score: (40.5% score 7/9 & 25% score 6/9), while 35 cases (17.5%) have moderate HCV activating score (15.5 % score 5/9 & 2 % score 4 / 9) and 10 cases (5%) have mild HCV activity score (4% score 3/9 and 1% score 2/9), while after treatment, find that significant improvement of HCV activity score (most of treated patients get improved HCV activity, 130 cases (65%) show mild HCV activity score (mostly 31.5% score 1/9; 20% score 3/9, 12.5% score 2/9 & 1% showed complete arrested activity score 0/9) while 22 cases (11%) show moderate HCV activity score (9% score 5/9 & 2% score 4/9) & few patients still have high HCV activity score 10% (are 9% score 6/9 & 1% score 7/9) (Figure 3).
To summarize the HCV activity score system before & after treatment we give the mean value of the activity score for all the patients, find that, the mean value of HCV activity score before treatment is high score 6.05/9, while after the treatment, there is significant marked improvement of HCV activity score lowered to the mild score 2.68/9.
b)Liver fibrosis Grading system: (normally no fibrosis) Liver fibrosis before the treatment find that (most of patients, 128 cases (64%) have moderate liver fibrosis G2, 34 cases (17%) have mild liver fibrosis G1 & few cases 4 (2%) have high liver fibrosis G3 as well as 8 cases (4%) have very high liver fibrosis G4, after the treatment, find that significant improvement of liver fibrosis Grading system as (most of treated patients) 124 cases (62%) became mild liver fibrosis G1 while 38 cases (19%) became moderate liver fibrosis G2 and 20 cases (10%) have very minimal liver fibrosis or no fibrosis as completely improved, but there is no cases with high nor very high liver fibrosis (Which indicate the success of the drug to treat the HCV & its complications. (Figure 4).
The new invented drug is a safe, effective and tolerable drug used as adjuvant treatment of HCV & its complications through the following mechanism of actions:
Mechanism of actions:
Mechanism of action of the main active consitnents4:
Squalene has a tritertenoid structure related to what we would like to call cyclosqualenoids.
Which is an open – chain 30 – carbon isoprenoid molecule, is cyclized (either directly, or via its metabolite, 2. 3 – oxidosqualene) to form tetracyclic or penta cyclic derivatives. The most familiar derivative which exist in nature is the proximate tatracyclic 30 – carbon molecule lanosterol5.
Squalene is an oily substance that easily absorbs oxygen and is presumed to help deliver oxygen throughout the body to the cell tissues that need it. This means that squalene can help enhance the quality of life, very useful for patients with heart diseases, diabetes, arthritis and hepatitis.
Squalene acts as antiviral agent, has been shown to have definite anti – inflammatory, anti – carcinogenic and antihepatitis.
The anti – inflammatory process and antiviral process are mechanistically related. Both pharmacological actions are based on their ability to block the de novo synthesis of two key anti inflammatory enzymes namely inducible oxide synthase (INOS) and inducible cyclooxygenase (COX-).
The mechanism of antihepatitis activity of squalene and its related compounds can be attributed to their absorbed oxygen content. These ozonized terpene hydrocarbons are act as antiviral or immunotherapeutic agents, which are useful in treating retroviral and flavi-viral infections where viral replication requires the transcription of viral RNA into DNA by viral reverse transcriptase enzyme. In some cases the antiviral agent prevents binding of certain viruses with their receptor on the cell surface6.
The therapeutic activity of silymarin is based on three sites or mechanisms of action:
a) It alters the structure of the outer cell membrane of the hepatocytes in such a way as to prevent penetration of the liver toxin and viruses into the interior of the cell.
b) It stimulates the action of nucleolar polymerase A, resulting in an increase in ribosomal protein synthesis and thus stimulates the generative ability of the liver and the formation of new hepatocytes.
c) It provides hepatocellular protection by stabilizing hepatic cell membranes. Other actions, include interruption of enterohepatic recirculation of toxins, stimulation of protein synthesis and regeneration of damaged hepatocytes as well as antioxidant activity.
III- Polyphenolic compounds mainly cynarin:
The commission E reported choleretic activity the British Herbal Pharmacopoeia reported hepatic action. Cynarin has demonstrated hepatoprotective and hepatostimulating properties.
IV- Biter compounds mainly secoiridoids:
Secoiridoids strengthen digestive function, especially within the stomach. By increasing stomach secretions, it hastens the breakdown of food. It also stimulates the appetite and increases bile production.
V- Sesquiterpene Lactones and Taraxacoside:
They have a significant cleansing action on the liver and stimulates bile production. It is also mildly gentle laxactives well as anticarcinogenic activity).
The other active constituents help to maintain the liver activity, strengthening digestive function, especially within the stomach.
Also it stimulate the appetite and increase bile production