WO2024040800A1

WO2024040800A1 - Use of reyanning in preparation of medicament for treating asymptomatic infection or mild infection by covid-19 omicron

Info

Publication number: WO2024040800A1
Application number: PCT/CN2022/138496
Authority: WO
Inventors: 杜成强
Original assignee: 清华德人西安幸福制药有限公司
Priority date: 2022-08-24
Filing date: 2022-12-12
Publication date: 2024-02-29

Abstract

The present invention relates to use of Reyanning in the preparation of a medicament for treating asymptomatic infection or mild infection by COVID-19 Omicron, which solves the problem that effective medicaments for Omicron variants in the prior art are very limited. According to the present invention, the cycle threshold (Ct value) of an open reading frame (ORF) or a nucleocapsid protein (N) gene of an asymptomatic infected individual or mildly infected individual can be increased within a short time. The Reyanning is prepared from water extracts of herba taraxaci, polygonum cuspidatum, herba patriniae and sculellaria barbata. The weight ratio of the herba taraxaci to the polygonum cuspidatum to the herba patriniae to the sculellaria barbata is (1-3):(1-3):(1-3):1. The Reyanning is prepared by the following preparation method: adding water to the above four medicinal materials for decoction twice, with 2 hours for the first time and 1 hour for the second time, filtering the decoctions, concentrating the filtrates under reduced pressure to a proper amount, combining the concentrated liquids, performing centrifugation and filtration, adding 1.5 g of steviosin and 0.5 g of ethylparaben, and heating to boiling, thus giving 1000 mL of the Reyanning.

Description

Application of Reyanning in the preparation of drugs for the treatment of asymptomatic or mild infections of COVID-19

Technical field

The present invention relates to the use of a traditional Chinese medicine composition, in particular to the use of Reyaning in the preparation of medicines for the treatment of asymptomatic or mild infections of COVID-19.

Background technique

COVID-19 has been spreading around the world for more than two years. As of June 2022, there have been more than 544 million confirmed cases of COVID-19 globally, including 6 million deaths. Many severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants, including Alpha, Beta, Gamma and Delta, have become an ongoing threat to global health. Since it was first reported in South Africa in November 2021, the emerging Omicron variant has rapidly swept the world. At present, the Omicron variant has become the main strain prevalent in the world. As of July 4, 2022, 193 countries have discovered and shared 4,501,325 Omicron genome sequences on the GISAID website. Omicron variants, including BA.1, BA.2, BA.3, BA.4, BA.5 and their descendant lineages, continue to evolve, leading to potential immune escape and higher transmission rates. As a result, Omicron was registered by the World Health Organization as a variant requiring immediate action (VOC).

Gene sequencing analysis by the Shanghai Center for Disease Control and Prevention laboratory showed that the new virus genome is mainly SARS-CoV-2BA.2.2 subline. Compared with the original strain of SARS-CoV-2, the Omicron variant has reduced pathogenicity but increased infectivity, leading to a sharp increase in the number of infected people and overloading of the medical system. Some previous studies have shown that the new coronavirus Omicron variant is mainly asymptomatic and mild infection, accompanied by upper respiratory tract symptoms such as cough, sputum, and fever. The "Novel Coronavirus Pneumonia Prevention and Control Plan (Eighth Edition)" defines asymptomatic infections as nucleic acid test positive (Ct value of N gene and ORF gene <35) and no clinical manifestations. According to the "Diagnosis and Treatment Plan for Novel Coronavirus Pneumonia (Trial Version 9)", mild disease is defined as pneumonia with mild clinical symptoms and no imaging evidence.

Antiviral drugs for SARS-CoV-2 can be divided into two categories: monoclonal antibodies against the Spike protein and small molecules that interfere with viral replication. Due to the high mutability of the Spike protein, most existing approved monoclonal antibodies have lost their neutralizing activity against SARS-CoV-2 Omicron. Currently, small molecule antiviral drugs targeting the conserved RdRp or Mpro of SARS-CoV-2 have been approved or emergency use authorization (EUA) and are in clinical trials. Remdesivir and monupivir are RdRp inhibitors of SARS-CoV-2, while nimaprevir/ritonavir (Paxlovid) targets Mpro. Previous clinical studies have shown that early use of these antiviral drugs reduces the risk of hospitalization or death in adults at high risk of mild to moderate Covid-19 who are not vaccinated and who are not hospitalized. However, these trials excluded patients who had been vaccinated against SARS-CoV-2; therefore, these findings may not apply to areas with high vaccination rates. Currently, there are very limited effective drugs against Omicron variants.

Contents of the invention

The purpose of the present invention is to provide an application of Reyaning in the preparation of medicines for the treatment of asymptomatic or mild infections of the new coronavirus Omicron, which overcomes the problem in the prior art that effective medicines for Omicron variants are very limited. The invention can increase the cycle threshold (Ct value) of the open reading frame (ORF) or nucleocapsid protein (N) gene in asymptomatic or mildly infected patients in a short period of time.

In order to achieve the above objects, the technical solutions adopted by the present invention are:

The application of Reyaning in the preparation of drugs for the treatment of asymptomatic infections or mild infections of COVID-19.

The application of Reyanning in the preparation of drugs that increase the cycle threshold (Ct) value of the open reading frame (ORF) or nucleocapsid protein (N) gene in asymptomatic infections or mild infections.

Reyanning is prepared from the water extracts of dandelion, Polygonum cuspidatum, Scutellaria barbata and Scutellaria barbata. The weight ratio of Dandelion, Polygonum cuspidatum, Scutellaria baicalensis and Scutellaria barbata is 1~3:1~3:1~3:1 .

Reyanning is prepared by the following preparation method: add the above four flavors and decoct them twice with water, the first time for 2 hours, the second time for 1 hour, filter the decoction, concentrate the filtrate under reduced pressure to an appropriate amount, combine the concentrates, and centrifuge. Filter, add 1.5g of stevia and 0.5g of ethyl hydroxyphenyl ester, heat to boiling, and make 1000ml.

The dosage of Reyanning is: 20ml of the traditional Chinese medicine composition mixture will be given from the first day, 4 times a day for 7 consecutive days.

Compared with the prior art, the present invention has the following advantages and effects:

1. The present invention can increase the cycle threshold (Ct value) of the open reading frame (ORF) or nucleocapsid protein (N) gene of asymptomatic infections or mild infections in a short period of time. It is a safe and effective treatment for asymptomatic and mild COVID-19 infection, which can accelerate virus clearance and promote disease recovery.

2. The Chinese medicinal composition of the present invention can be used to treat patients with asymptomatic COVID-19 infection or patients with mild COVID-19 infection. The cycle threshold (Ct value) of the open reading frame (ORF) or nucleocapsid protein (N) gene of SARS-CoV-2 was detected by RT-PCR and analyzed as semi-quantitative viral load. And through clinical trials, we can detect the nucleic acid negative conversion rate, nucleic acid negative conversion time and hospitalization time of asymptomatic or mildly infected patients, as well as the new symptoms and viral load of asymptomatic or mildly infected patients. To comprehensively evaluate the therapeutic effect of Reyanning Mixture in the treatment of patients with asymptomatic COVID-19 infection or mild COVID-19 infection. The results showed that RYN improved the nucleic acid negative conversion rate, shortened the nucleic acid negative conversion time and hospitalization time, reduced new symptoms in asymptomatic patients, and reduced the viral load. No serious adverse events were reported during the study. To sum up, it shows that the traditional Chinese medicine composition has significant therapeutic effect.

Description of drawings

The description and drawings that constitute a part of this application are used to provide a further understanding of the present invention. The illustrative embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the attached picture:

Figure 1 is a flow chart of the present invention;

Figure 2 is the calculation of negative conversion rate-KM analysis chart;

Figure 3 shows the negative conversion rate of nucleic acid. (A) Comparison of the negative conversion rate of nucleic acid within 3 days between the two groups. (B) Comparison of nucleic acid negative conversion rates within 7 days between the two groups;

Figure 4 is the discharge rate-KM analysis chart;

Figure 5 shows the new symptoms of asymptomatic infected persons during hospitalization;

Figure 6 shows the changes in SARS-CoV-2 viral load cycle threshold (Ct) during the observation process, (A) ORF gene Ct value. (B) N gene Ct value;

Figure 7 shows the changes in SARS-CoV-2 viral load cycle threshold (Ct) in subgroups of patients, (A-B) ORF/N gene Ct values in asymptomatic and mildly infected patients. (C-D) ORF/N gene Ct values in vaccinated and unvaccinated individuals.

Detailed ways

It should be noted that, as long as there is no conflict, the embodiments and features in the embodiments of this application can be combined with each other. The present invention will be described in detail below with reference to the accompanying drawings and embodiments.

The present invention includes the application of Reyaning in the preparation of medicines for treating asymptomatic or mild infections of COVID-19. In particular, it can improve the open reading frame (ORF) of asymptomatic or mild infections in a short period of time. or the cycle threshold (Ct value) of the nucleocapsid protein (N) gene. Reyanning is composed of four medicinal herbs: Dandelion (372 parts), Polygonum cuspidatum (372 parts), Beibeijiang (372 parts), and Scutellaria barbata (186 parts). Reyanning is prepared by the following preparation method: add the above four flavors and decoct them twice with water, the first time for 2 hours, the second time for 1 hour, filter the decoction, concentrate the filtrate under reduced pressure to an appropriate amount, combine the concentrates, and centrifuge. Filter, add 1.5g of stevia and 0.5g of ethyl hydroxyphenyl ester, heat to boiling, and make 1000ml.

Reyaning can be used to treat patients with asymptomatic or mild SARS-CoV-2 novel coronavirus infection. The cycle threshold (Ct value) of the open reading frame (ORF) or nucleocapsid protein (N) gene of SARS-CoV-2 was detected by RT-PCR and analyzed as semi-quantitative viral load. And through clinical trials, we can detect the nucleic acid negative conversion rate, nucleic acid negative conversion time and hospitalization time of asymptomatic or mildly infected patients, as well as the new symptoms and viral load of asymptomatic patients. To comprehensively evaluate the therapeutic effect of Reyanning Mixture (RYN) in the treatment of patients with asymptomatic or mild SARS-CoV-2 novel coronavirus infection. The results showed that RYN improved the nucleic acid negative conversion rate, shortened the nucleic acid negative conversion time and hospitalization time, reduced new symptoms in asymptomatic or mild patients, and reduced the viral load.

Experimental example:

1Research methods

1.1 Research Overview

This study adopted a prospective randomized controlled study method to recruit patients with COVID-19 asymptomatic or mild infection from the Fangcang Hospital of Shanghai New International Expo Center in China. The study protocol was designed in accordance with good clinical practice and the Declaration of Helsinki. The research protocol was approved by the Medical Ethics Committee of Jiujiang Hospital of Traditional Chinese Medicine (batch number: JJSZYYY20220403). The protocol has been registered on the Chinese Clinical Trial Registration website (www.chictr.org/cn/, ChiCTR2200060292). Written informed consent was obtained from all patients.

1.2 Research design

This study was a prospective, open-label, randomized controlled trial. Researchers enrolled subjects continuously. Serial numbers were prepared by a statistician not involved in the trial. According to the random number table generated by SPSS software, eligible patients were randomly divided into the intervention group (RYN+standard care) and the control group (standard care) in a 1:1 ratio. Treatment medications will not be concealed from investigators or patients. All participants received standard treatment (physical condition monitoring, antiviral, antibacterial, symptomatic treatment, and basic disease treatment) in accordance with the "New Coronavirus Pneumonia Diagnosis and Treatment Plan (Trial Version 9)".

Patients in the intervention group were given additional RYN mixture (20 ml, 4 times a day) from the first day of enrollment for 7 consecutive days. After 7 days of treatment, standard care was continued until discharge. Other concomitant medications should be carefully documented in the study. All patients were followed during hospitalization and for a maximum of 7 days after discharge within 7 days. The study was considered completed if the patient required transfer to a hospital for further treatment or discharge due to worsening condition.

Demographic, clinical, and epidemiological characteristics were investigated on the first day of enrollment. Eligible patients were studied by collecting data on clinical symptoms, comorbidities, concomitant medications, vaccination history, clinical outcomes, and adverse events.

Throat swab specimens were collected every day from the third day of admission for real-time polymerase chain reaction (RT-PCR) analysis until the nucleic acid turned negative. RT-PCR was used to detect the cycle threshold (Ct value) of the open reading frame (ORF) or nucleocapsid protein (N) gene of SARS-CoV-2 as a semi-quantitative viral load for analysis.

1.3 Research objects

Asymptomatic or mild patients will be recruited at the Shanghai New International Expo Center Fangcang Hospital from April 15 to May 12, 2022. The inclusion criteria are as follows: 1) Meet the diagnostic criteria for COVID-19 asymptomatic or mild infection; 2) Patients are aged 18 to 80 years old, regardless of gender; 3) Voluntarily provide informed consent.

Exclusion criteria include: 1) clinical classification of ordinary, severe, and critical; 2) patients with other severe primary respiratory diseases, such as lung cancer, severe bronchiectasis, and interstitial lung disease; 3) severe systemic diseases (such as malignant tumors, autoimmune diseases, blood, metabolic and endocrine diseases) and conditions involving important organs (heart, brain, liver, kidney), which may affect the evaluation of efficacy; 4) Pregnant women and lactating women; 5) Mental disorders ; 6) Participate in other clinical trials within 3 months; 7) Allergy or history of allergies to clinical trial drugs and conventional Western medicines.

1.4 Investigational Drugs

The test substance RYN is composed of dandelion, knotweed, soybean and barbata. RYN (batch number 220222) is produced by Tsinghua Deren Xi'an Xingfu Pharmaceutical Co., Ltd. This product was launched in 2005 and was approved by the State Food and Drug Administration (approval number: Z20050493). According to the "Chinese Pharmacopoeia" (2020 edition), 1000ml of RYN is taken from 372g dandelion, 372g Polygonum cuspidatum, 372g Scutellaria baicalensis, and 186g Scutellaria barbata. Use fingerprints for quality control and ingredient identification of medicinal materials. The results show that the main components of cyanin are chlorogenic acid, caffeic acid, resveratrol glycoside, luteolin and emodin.

1.5 Study endpoints

The main results of this test are the nucleic acid negative conversion time and the nucleic acid negative conversion rate on the 3rd and 7th days.

Secondary outcomes included length of hospital stay, new symptoms in asymptomatic infected patients, rate of disease progression during the observation period, and Ct value of RT-PCR detection.

The Ct value is inversely proportional to the viral load, that is, the higher the Ct value, the lower the mRNA expression. In samples where no Ct value was detected, the Ct value was automatically set to 45 for statistical purposes. Record all adverse events that may or may not be related to the study drug.

Nucleic acid negative conversion is defined as two consecutive N gene and ORF gene Ct values ≥35, and the nucleic acid detection interval is at least 24 hours.

The discharge criteria are as follows: (1) body temperature returns to normal for more than 3 days; (2) respiratory symptoms improve significantly; (3) nucleic acid conversion is negative.

1.6 Statistical methods

For continuous variables, independent samples t test and Mann-Whitney test were used to analyze normally distributed and non-normally distributed data, respectively. Normally distributed data are expressed as mean±standard deviation (mean±SD). Non-normally distributed data were expressed as the median with interquartile range (IQR). Categorical variables were expressed as frequencies and percentages (%), and analyzed using the chi-square test or Fisher's exact test. Time to event was expressed as median and 95% confidence interval (CI) and analyzed using Kaplan-Meier. Changes in SARS-CoV-2 viral load during the observation period were normalized using the local regression (loess) method. Statistical analysis was performed using R software (version 4.1.3). P<0.05 means the difference is statistically significant (two-sided test).

2 results

2.1 Baseline Statistics

Between April 14 and May 12, 2022, a total of 5,759 patients were reviewed, and 2,929 patients were excluded because they did not meet the inclusion criteria or refused to participate. Therefore, 2830 patients were randomly divided (1:1) into the intervention group (RYN+standard care) and the control group (standard care). In this trial, 65 patients in the intervention group dropped out and 27 in the control group dropped out. Finally, 1350 patients in the intervention group and 1388 patients in the control group were analyzed. The research flow chart is shown in Figure 1.

Table 1 summarizes baseline demographic and clinical characteristics. Among the 2830 patients, 2263 (80.0%) were asymptomatic and 567 (20.0%) were mildly infected. The median age of enrolled patients was 46 years old (IQR 33-56 years old), with 58.9% males and 41.1% females. The youngest patient is 18 years old and the oldest patient is 80 years old. Adults aged 18-44 have the highest proportion of patients. The most common symptoms on admission were cough (17.9%), sputum production (11.2%), fatigue (7.7%), fever (5.0%) and muscle aches (4.7%). Hypertension (12.4%) and diabetes (3.7%) were the most common comorbidities at baseline. 157 patients (5.5%) received antibiotics, 35 (1.2%) received antiviral therapy, and 2 (0.1%) received corticosteroids. The majority of patients were vaccinated, with 2104 (74.3%) receiving two or more doses of vaccine. Except for the history of close contact and antiviral treatment, there were no statistically significant differences in the baseline demographic and clinical characteristics of the two groups of patients (P>0.05).

Table 1 Baseline information and clinical characteristics.

2.2 Main results

The median time for nucleic acid to turn negative in the intervention group was 7 days (IQR: 4,10), and the median time for nucleic acid to turn negative in the control group was 8 days (IQR: 6,10.75). Kaplan-Meier analysis showed that patients receiving RYN treatment had a higher nucleic acid negative conversion rate (P<0.0001) (Figure 2). By the third day, 81 cases (6.0%) in the intervention group and 46 cases (3.3%) in the control group turned negative for nucleic acid. By day 7, 727 (53.9%) patients in the intervention group had negative nucleic acid tests, while only 520 (37.5%) patients in the control group had negative results.

The nucleic acid negative conversion rate of patients with asymptomatic infection and mild infection in the intervention group was significantly higher than that of the control group (P<0.05) (Figure 3).

2.3 Secondary results

There were no cases of disease progression during the observation period. The median length of stay in the intervention group was 8 days (IQR: 5,11) and in the control group was 9 days (IQR: 7,11). Kaplan-Meier analysis showed that patients treated with RYN had a higher discharge rate (P<0.0001) (Figure 4).

Among 2,263 patients who were asymptomatic upon admission, 325 in the intervention group and 452 in the control group developed clinical symptoms during the observation period. The proportion of new symptoms among patients in the intervention group, such as fever (2.4% vs. 4.5%, P = 0.008), cough (14.3% vs. 17.9%, P = 0.038), and expectoration (6.7% vs. 9.7%, P = 0.014), which were significantly lower than those of the control group (Figure 5).

The viral load Ct values of the SARS-CoV-2 ORF gene and N gene at each time point were drawn using the loess method to create a trend line. As shown in Figure 6, the patient's Ct value continued to increase and leveled off after the 9th day. After the Ct value reaches 40, the smooth curve becomes stable. The Ct value of the ORF/N gene in the RYN intervention group was higher than that in the control group (Figure 6A).

As shown in Figure 7, the Ct value of asymptomatic infections was higher than that of symptomatic infections. Among patients with asymptomatic infection, those who received RYN treatment had higher Ct values and reached nucleic acid negative criteria faster (Figure 7A,B). As can be seen from Figure 7C,D, the Ct value of vaccinated infected persons was higher than that of unvaccinated persons. In addition, RYN treatment promoted nucleic acid negative conversion, especially in unvaccinated patients.

2.4 Security analysis

During the observation process, a total of 303 single adverse events (AEs) were observed, including 150 cases in the intervention group and 153 cases in the control group, which were assessed as mild adverse events. The most common adverse events were diarrhea (2.2%), stomach pain (2.6%), anxiety (1.9%), and constipation (1.2%). Among all adverse events, diarrhea was assessed as possibly related to RYN treatment, while others were assessed as not related to the drug. There was no statistical difference in the incidence of AE between the two groups (Table 2).

Table 2 Adverse events

3 Discussion

In this prospective, open-label, randomized controlled trial, we provide clinical evidence of RYN in the treatment of SARS-CoV-2 COVID-19 infection, demonstrating that RYN is a safe treatment associated with rapid viral clearance and disease Recovery related. Among them, compared with standard care, RYN increased the nucleic acid negative conversion rate, shortened the negative conversion time and hospitalization time, reduced new symptoms in asymptomatic or mild patients, and lowered the viral load. For patients with asymptomatic or mild infection upon admission, RYN can inhibit the progression of asymptomatic infection to mild infection. Additionally, RYN has a favorable safety profile in COVID-19 patients, with no serious side effects reported. These findings suggest that RYN is a promising treatment for the Omicron variant of SARS-CoV-2.

From the data of this study, the overall vaccination rate among infected people reached 78%, and nearly half (45.6%) of the patients received booster shots. Combined with previous studies in blood samples and real-world studies, all the evidence suggests that Omicron produces immune evasion, whether through past infection or vaccination.

In summary, RYN is a safe and effective therapy for the treatment of asymptomatic and mild COVID-19 infection, which can accelerate virus clearance and promote disease recovery.

The above descriptions are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and changes. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention shall be included in the protection scope of the present invention.

Claims

The application of Reyaning in the preparation of drugs for the treatment of asymptomatic infections or mild infections of COVID-19.
The application of Redyanning in the preparation of medicines for treating asymptomatic or mild infections of COVID-19. The Redyanning is prepared from water extracts of dandelion, Polygonum cuspidatum, Beibeijiang and Scutellaria barbata.
The application of Reyaning according to claim 1 or 2 in the preparation of medicines for the treatment of asymptomatic infections or mild infections of COVID-19, characterized in that: the use of Reyaning in the preparation of drugs to improve the openness of asymptomatic infections The cycle threshold (Ct) value of the reading frame (ORF) or nucleocapsid protein (N) gene for pharmaceutical applications.
The application of Reyaning according to any one of claims 1 to 3 in the preparation of medicines for the treatment of asymptomatic or mild infections of COVID-19, characterized in that: the Reyaning is made of dandelion, Polygonum cuspidatum, It is prepared from the water extracts of dandelion and Scutellaria barbata, and the weight ratio of dandelion, Polygonum cuspidatum, Scutellaria baicalensis and Scutellaria barbata is 1 to 3:1 to 3:1 to 3:1.
The application of Redyanning according to claim 4 in the preparation of medicines for treating asymptomatic or mild infections of COVID-19, characterized in that: the Redyanning is prepared by the following preparation method: the above four flavors, Add water and cook twice, the first time for 2 hours and the second time for 1 hour. Filter the decoction and concentrate the filtrate under reduced pressure to an appropriate amount. Combine the concentrates, centrifuge, filter, and add 1.5g of stevia and 0.5g of ethyl hydroxyphenyl ester. g, heat to boiling, and make 1000ml.
The application of Redyanning according to claim 5 in the preparation of medicines for the treatment of asymptomatic or mild infections of COVID-19, characterized in that: the dosage of Redyanning is: a Chinese medicine combination is given from the first day 20ml of compound, 4 times a day for 7 days.
A method for treating asymptomatic or mild infection of COVID-19, which is characterized by comprising: administering an effective dose of Reyanin to a treatment subject.
The method according to claim 7, characterized in that the Reyaning is prepared from water extracts of Dandelion, Polygonum cuspidatum, Beibeijiang and Scutellaria barbata.
The method according to claim 7, characterized in that the dosage of Reyaning is: 20 ml of the traditional Chinese medicine composition mixture is administered from the first day, 4 times a day, for 7 consecutive days.
The method according to any one of claims 7 to 9, characterized in that the treatment of asymptomatic infection or mild infection of COVID-19 includes increasing the open reading frame of asymptomatic or mild infection or One or more of the cycle threshold Ct value of the nucleocapsid protein N gene, nucleic acid negative conversion time and hospitalization time, new symptoms in asymptomatic infections or mild infections, and accelerated virus clearance.
The method according to claims 8 to 8, characterized in that the Reyaning is prepared from water extracts of Dandelion, Polygonum cuspidatum, Scutellaria Scutellariae and Scutellaria Scutellariae, and The weight ratio is 1~3:1~3:1~3:1.
The method according to claim 8, characterized in that the Reyaning is prepared by the following preparation method: add water to decoct the above four flavors twice, the first time for 2 hours, the second time for 1 hour, and filter the decoction. Pass, concentrate the filtrate under reduced pressure to an appropriate amount, combine the concentrated solutions, centrifuge, filter, add 1.5g of stevia and 0.5g of ethyl hydroxyphenyl ester, heat to boiling, and make 1000ml.