diff --git a/.changeset/beige-results-flow.md b/.changeset/beige-results-flow.md
new file mode 100644
index 000000000000..3425c18594ad
--- /dev/null
+++ b/.changeset/beige-results-flow.md
@@ -0,0 +1,11 @@
+---
+"@cloudflare/vite-plugin": patch
+---
+
+Support Hyperdrive local connection strings from `.env` files
+
+You can now define your Hyperdrive local connection string in a `.env` file using the `CLOUDFLARE_HYPERDRIVE_LOCAL_CONNECTION_STRING_<BINDING_NAME>` variable.
+
+```sh
+CLOUDFLARE_HYPERDRIVE_LOCAL_CONNECTION_STRING_PROD_DB="postgres://user:password@127.0.0.1:5432/testdb"
+```
diff --git a/packages/vite-plugin-cloudflare/playground/bindings/.env b/packages/vite-plugin-cloudflare/playground/bindings/.env
new file mode 100644
index 000000000000..435034d90afa
--- /dev/null
+++ b/packages/vite-plugin-cloudflare/playground/bindings/.env
@@ -0,0 +1,2 @@
+# To test whether vite plugin could configure the hyperdrive binding properly
+CLOUDFLARE_HYPERDRIVE_LOCAL_CONNECTION_STRING_HYPERDRIVE="postgres://user:password@example.com:3456/testdb"
diff --git a/packages/vite-plugin-cloudflare/playground/bindings/__tests__/worker.spec.ts b/packages/vite-plugin-cloudflare/playground/bindings/__tests__/worker.spec.ts
index dd54c1f5c9a4..b3bc399c571d 100644
--- a/packages/vite-plugin-cloudflare/playground/bindings/__tests__/worker.spec.ts
+++ b/packages/vite-plugin-cloudflare/playground/bindings/__tests__/worker.spec.ts
@@ -25,3 +25,8 @@ test("ratelimit support", async () => {
 	const response = await getTextResponse("/rate-limit");
 	expect(response).toBe("Rate limit binding works: first: true, second: false");
 });
+
+test("hyperdrive support", async () => {
+	const response = await getTextResponse("/hyperdrive");
+	expect(response).toBe("Hyperdrive binding works");
+});
diff --git a/packages/vite-plugin-cloudflare/playground/bindings/src/index.ts b/packages/vite-plugin-cloudflare/playground/bindings/src/index.ts
index a4e5283ce1ea..4520a0cdebb5 100644
--- a/packages/vite-plugin-cloudflare/playground/bindings/src/index.ts
+++ b/packages/vite-plugin-cloudflare/playground/bindings/src/index.ts
@@ -61,6 +61,18 @@ export default {
 					}
 				);
 			}
+			case "/hyperdrive": {
+				if (
+					typeof env.HYPERDRIVE.connect !== "function" ||
+					typeof env.HYPERDRIVE.connectionString !== "string"
+				) {
+					return new Response("Hyperdrive binding is not configured properly", {
+						status: 500,
+					});
+				}
+
+				return new Response("Hyperdrive binding works");
+			}
 			case "/hello-world": {
 				const value = Math.floor(Date.now() * Math.random()).toString(36);
 				await env.HELLO_WORLD.set(value);
diff --git a/packages/vite-plugin-cloudflare/playground/bindings/worker-configuration.d.ts b/packages/vite-plugin-cloudflare/playground/bindings/worker-configuration.d.ts
index 5ff75b054ab0..69686c53bb66 100644
--- a/packages/vite-plugin-cloudflare/playground/bindings/worker-configuration.d.ts
+++ b/packages/vite-plugin-cloudflare/playground/bindings/worker-configuration.d.ts
@@ -7,6 +7,7 @@ declare namespace Cloudflare {
 		IMAGES: ImagesBinding;
 		WAE: AnalyticsEngineDataset;
 		RATE_LIMITER: RateLimit;
+		HYPERDRIVE: Hyperdrive;
 	}
 }
 interface Env extends Cloudflare.Env {}
diff --git a/packages/vite-plugin-cloudflare/playground/bindings/wrangler.jsonc b/packages/vite-plugin-cloudflare/playground/bindings/wrangler.jsonc
index e1f3f91354f8..ec364f8ce90d 100644
--- a/packages/vite-plugin-cloudflare/playground/bindings/wrangler.jsonc
+++ b/packages/vite-plugin-cloudflare/playground/bindings/wrangler.jsonc
@@ -23,6 +23,12 @@
 			"binding": "WAE",
 		},
 	],
+	"hyperdrive": [
+		{
+			"binding": "HYPERDRIVE",
+			"id": "test-hyperdrive-id",
+		},
+	],
 	"unsafe": {
 		"bindings": [
 			{
diff --git a/packages/vite-plugin-cloudflare/src/plugin-config.ts b/packages/vite-plugin-cloudflare/src/plugin-config.ts
index d57093164388..136c2200704f 100644
--- a/packages/vite-plugin-cloudflare/src/plugin-config.ts
+++ b/packages/vite-plugin-cloudflare/src/plugin-config.ts
@@ -112,6 +112,15 @@ export function resolvePluginConfig(
 		experimental: pluginConfig.experimental ?? {},
 	};
 	const root = userConfig.root ? path.resolve(userConfig.root) : process.cwd();
+	const prefixedEnv = vite.loadEnv(viteEnv.mode, root, [
+		"CLOUDFLARE_",
+		// TODO: Remove deprecated WRANGLER prefix support in next major version
+		"WRANGLER_HYPERDRIVE_LOCAL_CONNECTION_STRING_",
+	]);
+
+	// Merge the loaded env variables into process.env so that they are available to
+	// wrangler when it loads the worker configuration files.
+	Object.assign(process.env, prefixedEnv);
 
 	if (viteEnv.isPreview) {
 		return {
@@ -122,12 +131,7 @@ export function resolvePluginConfig(
 	}
 
 	const configPaths = new Set<string>();
-	const { CLOUDFLARE_ENV: cloudflareEnv } = vite.loadEnv(
-		viteEnv.mode,
-		root,
-		/* prefixes */ ""
-	);
-
+	const cloudflareEnv = prefixedEnv.CLOUDFLARE_ENV;
 	const entryWorkerConfigPath = getValidatedWranglerConfigPath(
 		root,
 		pluginConfig.configPath