A randomized, double blind, cross over study to evaluate the amino acid absorption after the intake of whey protein along with probiotic Bacillus coagulans SNZ 1969 in normal, healthy, adult, human subjects.
试验速览
- 阶段
- 不适用
- 状态
- 已完成
- 发起方
- 入组人数
- 30
- 试验地点
- 1
- 主要终点
- 1. Rate of amino acid absorption: To determine the rate (in minutes) of amino acid absorption into the bloodstream after a two-week pattern of whey protein supplementation with and without the addition of Bacillus coagulans SNZ 1969.
研究概览
简要总结
STUDY TITLE:
A randomized, double blind, cross over study to evaluate the amino acid absorption after the intake of whey protein along with probiotic Bacillus coagulans SNZ 1969 in normal, healthy, adult, human subjects.Project No. : 23-071 Protocol Version No. : 01
Date : 31 Oct 2023
Supersedes Version No. : NA Supersedes Date : NA
Contract Research OrganizationJeevan Scientific Technology Ltd. Plot No. 1&2, Sai Krupa Enclave Near Lanco Hills, Golconda Post Hyderabad 500 008, Telangana, India.
Tel No.: +91-40-67364700
SponsorSanzyme Biologics Private Limited, India.
Plot 13, 8/2/120/13/5, Road No. 2, Sagar Society, Sri Nagar Colony, Kamalapuri Colony, Banjara Hills, Hyderabad, Telangana 500034.
Phone no: 040 4858 9999.
CONFIDENTIALITY STATEMENTThe information contained in this protocol, is the proprietary information of the Sanzyme Biologics Private Limited, India and Jeevan Scientific Technology Ltd. (or under its control). This protocol was developed for Sanzyme Biologics Private Limited, India by Jeevan Scientific Technology Ltd. and should not be disclosed to third party without written authorization from the Sponsor with the exception of regulatory agencies and applicable Independent Ethics Committee, study volunteers, study staff and study audit personnel.
PROTOCOLSUMMARY
| Study Title |
A randomized, double blind, cross over study to evaluate the amino acid absorption after the intake of whey protein along with probiotic
Bacillus coagulans SNZ 1969 in normal, healthy, adult, human subjects.
|Study aim and Objectives
The purpose of this study is to examine the pharmacokinetic pattern of amino acid absorption after a two-week pattern of whey protein supplementation with and without the addition of Bacillus coagulans SNZ 1969. This study is designed to examine the impact of adding Bacillus coagulans SNZ 1969 to a 25-g dose of whey protein concentrate on post-prandial changes in blood amino acids concentrations.
1. Rate of amino acid absorption: To determine the rate (in minutes) of amino acid absorption into the bloodstream after a two-week pattern of whey protein supplementation with and without the addition of Bacillus coagulans SNZ 1969.
2. Extent of amino acid absorption: To determine the extent (concentration in millimoles/Liter) of amino acid absorption into the bloodstream after a two-week pattern of whey protein supplementation with and without the addition of Bacillus coagulans SNZ 1969.
3. To investigate the acne development after a two-week pattern of whey protein supplementation with and without the addition of Bacillus coagulans SNZ 1969.
|Study Design
A randomized, double blind, cross over study to evaluate the amino acid absorption after the intake of whey protein along with probiotic Bacillus coagulans SNZ 1969.
|Subjects
A total of 30 normal, healthy, adult, human subjects meeting the
inclusion and exclusion criteria will be enrolled in the study.
|Investigational Products
Treatment A (Experimental): Whey Protein
- Probiotic (n=15) 25 gram dose of whey protein concentrate with Bacillus coagulans SNZ 1969 2 Billion CFU Powder per serving
|Treatment B (Active Comparator): Whey Protein (n=15)
25 gram dose of whey protein concentrate
|Randomization
Subjects who meet the eligibility criteria will be randomly assigned to receive the study drugs according to one of the two dosing sequences: AB or BA.
|Study Visits
Supplementation Period 01: Subjects should visit the clinical facility on day 0 (For randomization and supply of treatments for 13 days) and day 14 (For administration of 14th dose, blood sample collection) of Supplementation period 01.
Supplementation Period 02: On day 34, Supply of treatments for supplementation period 02 and evaluation of acne development after completion of washout period of at least 21 days, Subjects should
| visit the clinical facility on day 48 (For administration of 14th dose, For blood sample collection) of | |
| Supplementation period 02. |
|Washout Period
A washout period of at least 21 days will be maintained between
consecutive Supplementation periods.
|Duration of the
study
Total expected duration of the study will be approximately 49 days (Including the washout period will be of at least 21 days).
|Methodology
The study will be conducted using a randomized, double-blind, crossover study design. 30 healthy adult human subjects between the ages of 18 - 30 years will be recruited to participate in this study.
Prior to beginning the study, all participants will sign an Ethics committee approved informed consent document and complete a healthy history questionnaire to determine study eligibility.
The study will be conducted in two supplementation periods with each span of two weeks and separated with a washout period of 21 days.
All the subjects will undergo a screening procedure comprising of clinical examination, recording of electrocardiogram and laboratory investigation of blood as well as urine which will be valid for 21 days from the day of screening. Radiological investigations (chest X-ray) will be repeated, if not done in the past 6 months or if clinically indicated at the time of screening. Subjects must be enrolled in the study only after providing written informed consent. Selection of subjects for the study will be done based on assessment against the inclusion and exclusion criteria.
On day 0, Clinical examination and vital signs (seated blood pressure, respiratory rate, radial pulse rate and body temperature), well-being and body mass index will be recorded at the time of visit on day 0.
The study participants will be randomly assigned to ingest either a single daily 25 grams dose of whey protein concentrate with Bacillus coagulans SNZ 1969 2 Billion CFU Powder (Treatment A) or a single daily 25 grams dose of whey protein concentrate (Treatment B). The sufficient quantities of either Treatment A or Treatment B will be supplied to study participants for 13 days supplementation (Day 1 to Day 13).
The study participants will be instructed to ingest the provided dose at same time for day 1 to day 13. The powder of either Treatment A or Treatment B will be mixed thoroughly with approximately 300 mL of water at ambient temperature and ingest between 08:00 to 10:00 hours at least 1 hour prior to the breakfast. All participants will be required to complete a supplementation log to document when the each dose of their assigned treatment was consumed.
| On day 14 and day 48, After consuming 13 consecutive doses, the study participants will | |
| report to the clinical facility between 06:00 to 08:00 hours, after observing | |
| an 8.00 to 10.00 hours fasting. |
Vital signs (seated blood pressure, respiratory rate, radial pulse rate and body temperature), well-being and body mass index will be recorded prior to ingestion of their 14th and final assigned supplementation dose.
The venous blood sample of 10 mL (2x5mL in sodium heparin vacutainer) will be collected at prior to ingest the 14th and final dose of their assigned supplement.
The study participants will be instructed to ingest the 14th or final dose of their final assigned supplementation dose. Upon ingestion of their final assigned supplementation dose, the subsequent venous blood samples of 10 mL each will be collected at 00.50 (30 minutes),
01.00 (60 minutes), 01.50 (90 minutes), 02.00 (120 minutes), 03.00
(180 minutes), and 04.00 (240 minutes) hour after ingestion.
The participants will be provided 200 mL of water to ingest after each blood collection.
The study participants will be discharged after 05.00 hours post ingest of powder. The clinical examination and vital signs during discharge of each visit may be started 120 minutes prior to the scheduled time of discharge of each subject, well-being of the subject will be confirmed at the time of actual discharge in each visit.
On day 34, Vital signs (seated blood pressure, respiratory rate, radial pulse rate and body temperature), well-being will be recorded.
Study participants will be provided an additional 13 days (Day 35 to 47) supply of the alternative treatment to begin after observing a three-week washout. After 13 days of supplementation, study participants will return to the clinical facility for their remaining testing visit. All subsequent study visits will be completed in a randomized fashion to minimize any order effects from testing.
Acne grading system will be done at baseline, end of 2 weeks of protein supplementation Period 01 (day 14) and prior to start of protein supplementation Period 01 (Day 34) and end of 2 weeks of protein supplementation Period 02 (day 48). To investigate If subject experience any acne development or that worsens during the study.
Acne grading will be evaluated by a dermatologist. Photographs will be taken at each time of acne grade assessment
| No. of | |
| Blood samples in supplementation period 01 |
07
|No. of Blood samples in supplementation period 02
07
|Total blood samples for both supplementation periods
14
|Volume of each sample
10 mL
| The below Acne grading system will be used to know the | |||||
| acne development. |
| Table: Grading severity of acne
|Grade
Severity
Clinical findings
|I
Mild
Open and closed comedones with few inflammatory papules and
pustules
|II
Moderate
Papules and pustules, mainly on face
|III
Moderately severe
Numerous papules and pustules, and occasional inflamed nodules, also on chest and back
|IV
Severe
Many large, painful nodules and
pustules
|Overview****of Study design
|Blood Sampling Schedule
The venous blood sample of 10 mL (2x5mL in sodium heparin vacutainer) will be collected at prior to ingest the 14th and final dose of their assigned supplement at each supplementation period.
The subsequent venous blood samples of 10 mL each will be collected at 00.50 (30 minutes), 01.00 (60 minutes), 01.50 (90
minutes), 02.00 (120 minutes), 03.00 (180 minutes), and 04.00 (240
| minutes) hour after | |
| ingestion in Sodium heparin vacutainer during each supplementation period. |
Total blood loss will not exceed 167.00 mL for each male subject and 176.00 mL for each female subject.
|Clinical Safety Measures
On day 0, Clinical examination and vital signs (seated blood pressure, respiratory rate, radial pulse rate and body temperature), well-being and body mass index will be recorded at the time of visit on day 0.
On day 14 and day 48, Vital signs (seated blood pressure, respiratory rate, radial pulse rate and body temperature), well-being and body mass index will be recorded prior to ingest of their 14th and final assigned supplementation dose.
The study participants will be discharged after 05.00 hours post ingest of powder. The clinical examination and vital signs (seated blood pressure, respiratory rate, radial pulse rate and body temperature) during discharge of each visit may be started 120 minutes prior to the scheduled time of discharge of each subject, well-being of the subject will be confirmed at the time of actual discharge in each visit.
On day 34, Vital signs (seated blood pressure, respiratory rate, radial pulse rate and body temperature), well-being will be recorded.
Clinical examination and measurement of vital signs may also be carried out at any time during the conduct of the study if the attending physician feels it necessary. In case of abnormality vital signs during pre-ingestion, medical opinion will be taken to dose the subject.
|Bioanalytical Methodology
Plasma samples will be assayed for concentration of different amino acids (1-Methyl-histidine, 3-Methyl- histidine, Alpha Aminoadipic acid, Alpha Amino butyric acid, Alanine, Anserine, Arginine, Argino succinic acid, Asparagine, Aspartic acid, Beta -Amino- isobutyric acid, Beta- Alanine, Carnosine, Citrulline, Cystathionine, Cystine, Ethanolamine, Gamma Amino isobutyric acid, Glutamine, Glutamic acid, Glycine, Homocitrulline, Homocystine, Histidine, Hydroxylysine, Hydroxyproline, Isoleucine, Leucine, Lysine, Methionine, Methionine sulfoxide, Norleucine, Norvaline, Ornithine, Phosphoethanolamine, Phenyl alanine, Proline, Phosphoserine, Sarcosine, Serine, Taurine, Threonine, Tryptophan, Tyrosine, Valine) using a LC-MS/MS in Dr. Lal Path Labs., National Reference Laboratory, B7 Road, Rohini Sector 18, Delhi 110085 (Near Astha Kunj Apartment & Sector 18 Metro Station,
Beside Petrol Pump, Block E)
|Pharmacokinetic Parameters
The following pharmacokinetic parameters will be estimated using Phoenix® WinNonlin® Version 8.4 or higher.
| AUC, Cmax and Tmax for different amino acids | |
| (1-Methyl-histidine, 3-Methyl- histidine, Alpha | |
| Aminoadipic acid, Alpha | |
| Amino butyric acid, Alanine, | |
| Anserine, Arginine, Argino succinic acid, Asparagine, Aspartic acid, Beta | |
| -Amino-isobutyric acid, Beta- Alanine, Carnosine, Citrulline, Cystathionine, | |
| Cystine, Ethanolamine, Gamma Amino | |
| isobutyric acid, Glutamine, Glutamic acid, Glycine, Homocitrulline, Homocystine, | |
| Histidine, Hydroxylysine, Hydroxyproline, Isoleucine, Leucine, Lysine, | |
| Methionine, Methionine sulfoxide, Norleucine, Norvaline, Ornithine, | |
| Phosphoethanolamine, Phenyl alanine, Proline, Phosphoserine, Sarcosine, Serine, Taurine, Threonine, Tryptophan, Tyrosine, Valine) |
Primary outcomes for this trial will be considered to be the area under the curve (AUC) data for the measured amino acids.
Secondary outcomes will be considered to the maximum concentrations (Cmax) identified for the measured amino acids.
|Study assessment
For all dependent measures, descriptive statistics will be presented as mean ± standard deviations. Before any statistical tests, the normality will be assessed for all dependent variables. All non- normal data will be log transformed and then analyzed using both parametric and non-parametric approaches. All reported p values will be computed using parametric approaches. Paired sample t-tests will be used to determine between-group differences for the AUC, Cmax, and Tmax values for all individually measured amino acids
Statistical analysis will be performed using SAS®, Version 9.4 or higher.
The appropriate statistical approach will be followed for analysis of acne development data.
研究设计
- 研究类型
- Interventional
- 分配方式
- Randomized
- 盲法
- Participant and Investigator Blinded
入排标准
- 年龄范围
- 18.00 Year(s) 至 30.00 Year(s)（—）
- 性别
- All
入选标准
- •Healthy human subjects between 18-30 years of age (including both) and weight and BMI 18.5 to 30.0 kg/m² (including both) with body fat <30%.
- •Acceptable findings during registration and screening including, medical history, physical examination, laboratory evaluations, 12- lead ECG and Chest X-Ray (postero-anterior view).
- •Non-smokers and Non-alcoholics.
- •Completing at least 30 minutes of moderate exercise three days per week.
- •Values within normal ranges of laboratory parameters upon evaluation by the Investigator or Physician for any of the following tests.
- •Subjects able to communicate effectively.
- •Subjects willing to give written informed consent and adhere to all the requirements of this protocol.
- •Subject willing to abstain from all kinds of caffeine/xanthine containing foods or grapefruit or grapefruit juice from 72.00 hours prior to visit until the last post dose blood sample collection in each supplementation period.
- •Female subjects: Postmenopausal for at least 1 year or Surgically sterile (bilateral tubal ligation, bilateral oophorectomy or hysterectomy has been performed on the subject) or Female of childbearing potential practicing an acceptable method of birth control for the duration of the study at least two weeks thereafter as judged by the investigator(s), such as condoms, foams, jellies, diaphragm, and intrauterine device (IUD) or abstinence.
排除标准
- •Volunteers having history of contraindication or hypersensitivity (e.g., anaphylaxis) to whey protein products.
- •A history of asthma, urticaria, or other allergic-type reactions after taking aspirin or other NSAIDs.
- •A history of seizures, diabetes, migraine, hypertension, cardiovascular, pulmonary, neurological or psychiatric disease/disorder, dermatological, endocrine, eye disorders, immunological, hepatic, renal, hematopoietic, metabolic, gastrointestinal, ongoing infectious diseases, or any other significant abnormality as evidenced by medical history and physical examination or according to the opinion of the physician.
- •History or presence of gastrointestinal (GI) inflammation, bleeding, ulceration, and perforation of the stomach, small intestine, or large intestine.
- •History or evidence of exfoliative dermatitis, Stevens.
- •Johnson syndrome (SJS), and toxic epidermal necrolysis (TEN).
- •Any known enzyme inducing or inhibiting drug taken within 30 days before the study.
- •Participants who are determined to not be weight stable defined as measured BMI deviating by 2% or more.
- •Participation in a drug research study within 90 days prior to intake of first supplementation.
- •Blood loss or whole blood donation within 90 days prior to intake of first supplementation.
- •Consumption of high caffeine (more than 5 cups of coffee or tea/day).
- •History of addiction to any recreational drug or drug dependence.
- •An unusual or abnormal diet, for whatever reason within 48.00 hours prior to visit of each supplementation period, e.g., fasting due to religious reasons.
- •History of dehydration from diarrhea, vomiting or any other reason within a period of 72.00 hours prior to intake of first supplementation
- •Positive results for drugs of abuse (benzodiazepines, cocaine, opioids, amphetamines, cannabinoids and barbiturates) in urine during the each visit of the study.
- •Positive results for alcohol consumption tests during the study admission of each visit.
- •History of pre-existing bleeding disorder.
- •Difficulty with donating blood.
- •Systolic blood pressure less than 100 mm Hg or more than 140 mm Hg.
- •Diastolic blood pressure less than 60 mm Hg or more than 90 mm Hg.
- •Pulse rate less than 60 beats/minute or more than 100 beats/minute.
- •Use of any prescribed medication or OTC medicinal products including vitamins and herbal medicinal products during last two weeks preceding the first dosing.
- •Participants who do not or are not willing to abstain from exercise for 12.00 hours prior to each visit.
- •Female volunteer who is pregnant, currently breast-feeding.
- •Female subjects demonstrating a positive pregnancy test.
- •Female volunteer who has used implanted or injected hormonal contraceptives anytime during the 6 months prior to study or used hormonal contraceptives within 14 days before first dosing.
结局指标
主要结局
1. Rate of amino acid absorption: To determine the rate (in minutes) of amino acid absorption into the bloodstream after a two-week pattern of whey protein supplementation with and without the addition of Bacillus coagulans SNZ 1969.
时间窗: At the end of 2 weeks of supplementation in Period 1 and period 2 respectively
2. Extent of amino acid absorption: To determine the extent (concentration in millimoles/Liter) of amino acid absorption into the bloodstream after a two-week pattern of whey protein supplementation with and without the addition of Bacillus coagulans SNZ 1969.
时间窗: At the end of 2 weeks of supplementation in Period 1 and period 2 respectively
次要结局
- To investigate the acne development after a two-week pattern of whey protein supplementation with and without the addition of Bacillus coagulans SNZ 1969.(Baseline and end of supplementation for Period I and II respectively)
研究者
Dr Mallu Maheswara Reddy
Jeevan Scientific Technology Limited